Down-regulating gene expression in insect pests

ABSTRACT

The present invention relates to genetic control of infestation by insect pest species, particularly prevention and/or control of pest infestation of plants, using interfering ribonucleic acid (RNA) molecules. Compositions and combinations containing the interfering RNA molecules of the invention for use in topical applications, for example in the form of insecticides.

RELATED APPLICATIONS

This application is a divisional application of application Ser. No.13/462,636, filed May 2, 2012, which claims was a continuation-in-partof PCT/EP2012/057332, filed Apr. 20, 2012, which claims the benefitunder 35 U.S.C. § 119(e) of U.S. provisional application 61/477,371filed Apr. 20, 2011, and U.S. provisional application 61/508,826 filedJul. 18, 2011, each of which is incorporated by reference in itsentirety herein.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted in ASCII format via EFS-Web and is hereby incorporated byreference in its entirety.

FIELD OF THE INVENTION

The present invention relates generally to genetic control ofinfestation by insect pest species, particularly prevention and/orcontrol of pest infestation of plants. More specifically, the inventionrelates to down-regulation of expression of target genes in insect pestspecies by interfering ribonucleic acid (RNA) molecules. Compositionsand combinations containing the interfering RNA molecules of theinvention for use in topical applications, for example in the form ofinsecticides, are also provided.

BACKGROUND TO THE INVENTION

There exists an abundance of insect pest species that can infect orinfest a wide variety of environments and host organisms. Insect pestsinclude a variety of species from the insect Orders Hemiptera (truebugs), Coleoptera (beetles), Siphonaptera (fleas), Dichyoptera(cockroaches and mantids), Lepidoptera (moths and butterflies),Orthoptera (e.g. grasshoppers) and Diptera (true flies). Pestinfestation can lead to significant damage. Insect pests that infestplant species are particularly problematic in agriculture as they cancause serious damage to crops and significantly reduce plant yields. Awide variety of different types of plant are susceptible to pestinfestation including commercial crops such as rice, cotton, soybean,potato and corn.

Traditionally, infestation with insect pests has been prevented orcontrolled through the use of chemical pesticides. However, thesechemicals are not always suitable for use in the treatment of crops asthey can be toxic to other species and can cause significantenvironmental damage. Over more recent decades, researchers havedeveloped more environmentally-friendly methods of controlling pestinfestation. For example, microorganisms such as Bacillus thuringiensisbacteria that naturally express proteins toxic to insect pests have beenused. Scientists have also isolated the genes encoding theseinsecticidal proteins and used them to generate transgenic cropsresistant to insect pests e.g. corn and cotton plants geneticallyengineered to produce proteins of the Cry family.

Although bacterial toxins have been highly successful in controllingcertain types of pest, they are not effective against all pest species.Researchers have therefore looked for other more targeted approaches topest control and in particular to RNA interference or ‘gene silencing’as a means to control pests at the genetic level.

RNA interference or ‘RNAi’ is a process whereby the expression of genesin the context of a cell or whole organism is down-regulated in asequence-specific manner. RNAi is now a well-established technique inthe art for inhibiting or down-regulating gene expression in a widevariety of organisms including pest organisms such as fungi, nematodesand insects. Furthermore, previous studies have shown thatdown-regulation of target genes in insect pest species can be used as ameans to control pest infestation.

WO2007/074405 describes methods of inhibiting expression of target genesin invertebrate pests including Colorado potato beetle. Furthermore,WO2009/091864 describes compositions and methods for the suppression oftarget genes from insect pest species including pests from the Lygusgenus.

Although the use of RNAi for down-regulating gene expression in pestspecies is known in the art, the success of this technique for use as apest control measure depends on selection of the most appropriate targetgenes, namely those wherein loss of function results in significantdisruption of an essential biological process and/or death of theorganism. The present invention is thus directed towards thedown-regulation of particular target genes in insect pests as a means toachieve more effective prevention and/or control of insect pestinfestation, particularly of plants.

SUMMARY OF THE INVENTION

The current inventors sought to identify improved means for preventingand/or controlling insect pest infestation using genetic approaches. Inparticular, they investigated the use of RNAi to down-regulate genes insuch a way as to impair the ability of the insect pest to survive, grow,colonize specific environments and/or infest host organisms and thuslimit the damage caused by the pest. Therefore, in accordance with oneaspect of the invention, there is provided an interfering ribonucleicacid (RNA or double stranded RNA) that functions upon uptake by aninsect pest species to down-regulate expression of a target gene in saidinsect pest,

wherein the RNA comprises at least one silencing element wherein thesilencing element is a region of double-stranded RNA comprising annealedcomplementary strands, one strand of which comprises or consists of asequence of nucleotides which is at least partially complementary to atarget nucleotide sequence within the target gene, and wherein thetarget gene(i) is selected from the group of genes having a nucleotide sequencecomprising any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13,14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62,19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78,143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135,226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153,246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389,or the complement thereof, or having a nucleotide sequence that, whenthe two sequences are optimally aligned and compared, is at least 75%,preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, or(ii) is selected from the group of genes having a nucleotide sequenceconsisting of any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189,27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6,35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50,13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132,214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126,135, 226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234to 237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152,153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159,258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167,166, 270 to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200,201, 314 to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387,388, 389, or the complement thereof, or(iii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 110, 125, 150, 175, 200,225, 250, 300, 350, 400, 450, 500, 550, 600, 700, 800, 900 1000, 1100,1200, 1300, 1400, 1500, 2000 or 3000 contiguous nucleotides of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, or having a nucleotide sequence that, when saidgene comprising said fragment is optimally aligned and compared with anyof SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389,said nucleotide sequence is at least 75% preferably at least 80%, 85%,90%, 95%, 98% or 99% identical to any of SEQ ID NOs 1, 174, 404, 180,188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3,4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46,141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71to 74, 25, 26, 75 to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128,149, 184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150,151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157,254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164,266 to 269, 165, 167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172,173, 278 to 281, 200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309,318 to 321, 386, 387, 388, 389, or the complement thereof, or(iv) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 110, 125, 150, 175, 200,225, 250, 300, 350, 400, 450, 500, 550, 600, 700, 800, 900 1000, 1100,1200, 1300, 1400, 1500, 2000 or 3000 contiguous nucleotides of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, and wherein when said fragment is optimallyaligned and compared with the corresponding fragment in any of SEQ IDNOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15,204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66,21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142,176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122,144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124,133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229,127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302 to305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to 249,154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160,161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273,168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to 317,186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, saidnucleotide sequence of said fragment is at least 75% preferably at least80%, 85%, 90%, 95%, 98% or 99% identical to said corresponding fragmentof any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30,282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38,140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20,63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143,121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301,145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237,302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, or(v) is an insect pest orthologue of a gene having a nucleotide sequencecomprising any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13,14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62,19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78,143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135,226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153,246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389,or the complement thereof, wherein the two orthologous genes are similarin sequence to such a degree that when the two genes are optimallyaligned and compared, the orthologue has a sequence that is at least 75%preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to any ofthe sequences represented by SEQ ID NOs 1, 174, 404, 180, 188, 2, 175,181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34,139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12,47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17,18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25,26, 75 to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293,123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225,147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184,137, 185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257,158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269,165, 167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to281, 200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309, 318 to 321,386, 387, 388, 389, or(vi) is selected from the group of genes having a nucleotide sequenceencoding an amino acid sequence that, when the two amino acid sequencesare optimally aligned and compared, is at least 70% preferably at least75%, 80%, 85%, 90%, 95%, 98% or 99% identical to the amino acid sequenceencoded by any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13,14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62,19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78,143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135,226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153,246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389.

In a particular aspect of the invention, interfering RNA molecules ofthe current invention comprise at least one double-stranded region,typically the silencing element of the interfering RNA, comprising asense RNA strand annealed by complementary basepairing to an antisenseRNA strand wherein the sense strand of the dsRNA molecule comprises asequence of nucleotides complementary to a sequence of nucleotideslocated within the RNA transcript of the target gene.

In one embodiment, the present invention relates to an interferingribonucleic acid (RNA or double stranded RNA) that functions upon uptakeby an insect pest species to down-regulate expression of a target genein said insect pest, wherein the RNA comprises at least one silencingelement wherein the silencing element is a region of double-stranded RNAcomprising annealed complementary strands, one strand of which comprisesor consists of a sequence of nucleotides which is at least partiallycomplementary to a target nucleotide sequence within the target gene,and wherein the target gene

(i) is selected from the group of genes having a nucleotide sequencecomprising any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27to 30, 282 to 285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177,183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179,210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146,125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to233 or the complement thereof, or having a nucleotide sequence so that,when the two sequences are optimally aligned and compared, is at least75% preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to anyof SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, or thecomplement thereof, or(ii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, or thecomplement thereof, or having a nucleotide sequence so that, when saidgene comprising said fragment is optimally aligned and compared with anyof SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, saidnucleotide sequence is at least 75% preferably at least 80%, 85%, 90%,95%, 98% or 99% identical to any of SEQ ID NOs 1, 174, 404, 180, 188, 2,175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 121, 142,176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122,144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124,133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229,127, 148, 136, 230 to 233, or the complement thereof, or(iii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, or thecomplement thereof, and wherein when said fragment is optimally alignedand compared with the corresponding fragment in any of SEQ ID NOs 1,174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297,310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289,298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123,132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147,126, 135, 226 to 229, 127, 148, 136, 230 to 233, said nucleotide sequence of said fragment is at least 75% preferablyat least 80%, 85%, 90%, 95%, 98% or 99% identical to said correspondingfragment of any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189,27 to 30, 282 to 285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130,177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131,179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221,146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230to 233, or the complement thereof, or(iv) is an insect pest orthologue of a gene having a nucleotide sequencecomprising any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27to 30, 282 to 285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177,183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179,210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146,125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to233, or the complement thereof, wherein the two orthologous genes aresimilar in sequence to such a degree that when the two genes areoptimally aligned and compared, the orthologue has a sequence that is atleast 75% preferably at least 80%, 85%, 90%, 95%, 98% or 99% identicalto any of the sequences represented by SEQ ID NOs 1, 174, 404, 180, 188,2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233 or(v) is selected from the group of genes having a nucleotide sequenceencoding an amino acid sequence that, when the two amino acid sequencesare optimally aligned and compared, is at least 85% preferably at least90%, 95%, 98% or 99% identical to the amino acid sequence encoded by anyof SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233.

These target genes encode proteins within the troponin/myofilamentcomplex.

In a further embodiment, the present invention relates to an interferingribonucleic acid (RNA or double stranded RNA) that functions upon uptakeby an insect pest species to down-regulate expression of a target genein said insect pest, wherein the RNA comprises at least one silencingelement wherein the silencing element is a region of double-stranded RNAcomprising annealed complementary strands, one strand of which comprisesor consists of a sequence of nucleotides which is at least partiallycomplementary to a target nucleotide sequence within the target gene,and wherein the target gene

(i) is selected from the group of genes having a nucleotide sequencecomprising any of SEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 150, 151, 242 to 245, 152, 153, 246 to 249,154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160,161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273,or the complement thereof, or having a nucleotide sequence so that, whenthe two sequences are optimally aligned and compared, is at least 75%preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to any ofSEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9,10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205,55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to70, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253,156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163,162, 164, 266 to 269, 165, 167, 166, 270 to 273, or the complementthereof, or(ii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9,10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205,55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to70, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253,156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163,162, 164, 266 to 269, 165, 167, 166, 270 to 273, or the complementthereof, or having a nucleotide sequence so that, when said genecomprising said fragment is optimally aligned and compared with any ofSEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9,10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205,55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to70, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253,156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163,162, 164, 266 to 269, 165, 167, 166, 270 to 273, said nucleotidesequence is at least 75% preferably at least 80%, 85%, 90%, 95%, 98% or99% identical to any of SEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38,140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20,63 to 66, 21, 22, 67 to 70, 150, 151, 242 to 245, 152, 153, 246 to 249,154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160,161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273,or the complement thereof, or(iii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9,10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205,55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to70, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253,156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163,162, 164, 266 to 269, 165, 167, 166, 270 to 273, or the complementthereof, and wherein when said fragment is optimally aligned andcompared with the corresponding fragment in any of SEQ ID NOs. 3, 4, 31to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141,11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 150, 151, 242to 245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to269, 165, 167, 166, 270 to 273, said nucleotide sequence of saidfragment is at least 75% preferably at least 80%, 85%, 90%, 95%, 98% or99% identical to said corresponding fragment of any of SEQ ID NOs. 3, 4,31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46,141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 150, 151,242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to269, 165, 167, 166, 270 to 273, or the complement thereof, or(iv) is an insect pest orthologue of a gene having a nucleotide sequencecomprising any of SEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 150, 151, 242 to 245, 152, 153, 246 to 249,154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160,161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273,or the complement thereof, wherein the two orthologous genes are similarin sequence to such a degree that when the two genes are optimallyaligned and compared, the orthologue has a sequence that is at least 75%preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to any ofthe sequences represented by SEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13,14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62,19, 20, 63 to 66, 21, 22, 67 to 70, 150, 151, 242 to 245, 152, 153, 246to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270to 273, or(v) is selected from the group of genes having a nucleotide sequenceencoding an amino acid sequence that, when the two amino acid sequencesare optimally aligned and compared, is at least 85% preferably at least90%, 95%, 98% or 99% identical to the amino acid sequence encoded by anyof SEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42,9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16,205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22,67 to 70, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265,163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273.

These target genes encode insect ribosomal proteins.

In certain embodiments, the present invention relates to an interferingRNA molecule which comprises at least one double-stranded region,typically the silencing element of the interfering RNA molecule,comprising a sense RNA strand annealed by complementary basepairing toan antisense RNA strand wherein the sense strand of the dsRNA moleculecomprises a sequence of at least 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 110, 125, 150,175, 200, 225, 250, 300, 350, 400, 450, 500, 550, 600, 700, 800, 9001000, 1100, 1200, 1300, 1400, 1500, 2000 or 3000 contiguous nucleotides,that is at least 75% preferably at least 80%, 85%, 90%, 95%, 98%, 99% or100% complementary to a sequence of nucleotides located within the RNAtranscript of a target gene from the troponin/myofilament complex.

In one embodiment, the target gene encodes an insect wings up A(troponin I) protein (e.g. an insect orthologue of the CG7178 Dmprotein), said target gene being represented by SEQ ID NOs 1, 2, 174,404, 175, 180, 181, 188 and 189. In a preferred embodiment, the insectorthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100% aminoacid sequence identity to one or more of SEQ ID NOs. 79, 349, 405, 352or 356.

In one embodiment, the target gene encodes an upheld protein (e.g. aninsect orthologue of the CG7107 Dm protein), said target gene beingrepresented by SEQ ID NOs 121, 130, 142, 143, 176, 177, 182 and 183. Ina preferred embodiment, the insect orthologue has at least 85%, 90%,92%, 94%, 96%, 98%, 99% or 100% amino acid sequence identity to one ormore of SEQ ID NOs. 330, 350 or 353.

In one embodiment, the target gene encodes the tropomyosin 1 protein(e.g. an insect orthologue of the CG4898 Dm protein), or the tropomyosin2 protein (e.g. an insect orthologue of the CG4843 Dm protein), saidtarget gene being represented by SEQ ID NOs 123 and 132. In a preferredembodiment, the insect orthologue has at least 85%, 90%, 92%, 94%, 96%,98%, 99% or 100% amino acid sequence identity to SEQ ID NO. 332.

In one embodiment, the target gene encodes the myosin heavy chain (e.g.an insect orthologue of the CG17927 Dm protein), said target gene beingrepresented by SEQ ID NOs 122, 131, 144, 145, 178 and 179. In apreferred embodiment, the insect orthologue has at least 85%, 90%, 92%,94%, 96%, 98%, 99% or 100% amino acid sequence identity to one or moreof SEQ ID NOs. 331 or 351.

In one embodiment, the target gene encodes the myosin light chaincytoplasmic protein (e.g. an insect orthologue of the CG3201 Dmprotein), said target gene being represented by SEQ ID NOs 124 and 133.In a preferred embodiment, the insect orthologue has at least 85%, 90%,92%, 94%, 96%, 98%, 99% or 100% amino acid sequence identity to SEQ IDNO. 333.

In one embodiment, the target gene encodes the spaghetti squash protein(e.g. an insect orthologue of the CG3595 Dm protein), said target genebeing represented by SEQ ID NOs 125 and 134. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% identity to SEQ ID NO. 334.

In one embodiment, the target gene encodes the zipper protein (e.g. aninsect orthologue of the CG15792 Dm protein), said target gene beingrepresented by SEQ ID NOs 126 and 135. In a preferred embodiment, theinsect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100%identity to SEQ ID NO. 335.

In one embodiment, the target gene encodes the troponin C (e.g. aninsect orthologue of the CG2981, CG7930, CG9073, CG6514, CG12408,CG9073, CG7930, CG2981, CG12408 or CG6514 Dm protein), said target genebeing represented by SEQ ID NOs 127 and 136, or 128 and 137, or 184 and185. In a preferred embodiment, the insect orthologue has at least 85%,90%, 92%, 94%, 96%, 98%, 99% or 100% amino acid sequence identity to oneor more of SEQ ID NOs. 336, 337 and 354.

According to another embodiment the present invention relates to aninterfering RNA molecule which comprises at least one double-strandedregion, typically the silencing element of the interfering RNA molecule,comprising a sense RNA strand annealed by complementary basepairing toan antisense RNA strand wherein the sense strand of the dsRNA moleculecomprises a sequence of at least 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 110, 125, 150,175, 200, 225, 250, 300, 350, 400, 450, 500, 550, 600, 700, 800, 9001000, 1100, 1200, 1300, 1400, 1500, 2000 or 3000 contiguous nucleotides,that is at least 75% preferably at least 80%, 85%, 90%, 95%, 98%, 99% or100% complementary to a sequence of nucleotides located within the RNAtranscript of a target gene that encodes an insect ribosomal protein.

In one embodiment, the target gene encodes ribosomal protein S3A (e.g.an insect orthologue of the CG2168 Dm protein), said target gene beingrepresented by SEQ ID NOs 11, 12 and 141. In a preferred embodiment, theinsect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100%amino acid sequence identity to one or both of SEQ ID NO. 84 or 328.

In one embodiment, the target gene encodes the ribosomal protein LP1(e.g. an insect orthologue of the CG4087 Dm protein), said target genebeing represented by SEQ ID NO 3 and 4. In a preferred embodiment, theinsect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100%amino acid sequence identity to SEQ ID NO.80.

In one embodiment, the target gene encodes the ribosomal protein S3(e.g. an insect orthologue of the CG6779 Dm protein), said target genebeing represented by SEQ ID NOs 7 and 8. In a preferred embodiment, theinsect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100%amino acid sequence identity to SEQ ID NO.82.

In one embodiment, the target gene encodes the ribosomal protein L10Ab(e.g. an insect orthologue of the CG7283 Dm protein) represented by SEQID NOs 9 and 10. In a preferred embodiment, the insect orthologue has atleast 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100% amino acid sequenceidentity to SEQ ID NO. 83.

In one embodiment, the target gene encodes the ribosomal protein S18(e.g. an insect orthologue of the CG8900 Dm protein), said target genebeing represented by SEQ ID NO 13 and 14. In a preferred embodiment, theinsect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100%amino acid sequence identity to SEQ ID NO.85.

In one embodiment, the target gene encodes the ribosomal protein L4(e.g. an insect orthologue of the CG5502 Dm protein), said target generepresented by SEQ ID NO 5 and 6. In a preferred embodiment, the insectorthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100% aminoacid sequence identity to SEQ ID NO.81.

In one embodiment, the target gene encodes the ribosomal protein S27(e.g. an insect orthologue of the CG10423 Dm protein), said target genebeing represented by SEQ ID NO 15 and 16, 204 and 205. In a preferredembodiment, the insect orthologue has at least 85%, 90%, 92%, 94%, 96%,98%, 99% or 100% amino acid sequence identity to one or both of SEQ IDNOs.86 and 359.

In one embodiment, the target gene encodes the ribosomal protein L6(e.g. an insect orthologue of the CG11522 Dm protein), said target genebeing represented by SEQ ID NO 17 and 18. In a preferred embodiment, theinsect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100%amino acid sequence identity to SEQ ID NO. 87.

In one embodiment, the target gene encodes the ribosomal protein S13(e.g. an insect orthologue of the CG13389 Dm protein), said target genebeing represented by SEQ ID NO 19 and 20. In a preferred embodiment, theinsect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100%amino acid sequence identity to SEQ ID NO. 88.

In one embodiment, the target gene encodes the ribosomal protein L12(e.g. an insect orthologue of the CG3195 Dm protein), said target genebeing represented by SEQ ID NOs 21 and 22. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO.89.

In one embodiment, the target gene encodes the ribosomal protein L26(e.g. an insect orthologue of the CG6846 Dm protein), said target genebeing represented by SEQ ID NOs 158 and 159. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO. 343.

In one embodiment, the target gene encodes the ribosomal protein L21(e.g. an insect orthologue of the CG12775 Dm protein), said target genebeing represented by SEQ ID NO 165, 166 and 167. In a preferredembodiment, the insect orthologue has at least 85%, 90%, 92%, 94%, 96%,98%, 99% or 100% amino acid sequence identity to SEQ ID NOs 347 and 348.

In one embodiment, the target gene encodes the ribosomal protein S12(e.g. an insect orthologue of the CG11271 Dm protein), said target genebeing represented by SEQ ID NOs 156 and 157. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO. 342.

In one embodiment, the target gene encodes the ribosomal protein S28b(e.g. an insect orthologue of the CG2998 Dm protein), said target genebeing represented by SEQ ID NOs 160 and 161. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO. 344.

In one embodiment, the target gene encodes the ribosomal protein L13(e.g. an insect orthologue of the CG4651 Dm protein), said target genebeing represented by SEQ ID NOs. 154 and 155. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO. 341.

In one embodiment, the target gene encodes the ribosomal protein L10(e.g. an insect orthologue of the CG17521 Dm protein), said target genebeing represented by SEQ ID NOs. 163 and 164. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO. 345.

In one embodiment, the target gene encodes the ribosomal protein L5(e.g. an insect orthologue of the CG17489 Dm protein), said target genebeing represented by SEQ ID NOs. 152 and 153. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO. 340.

In one embodiment, the target gene encodes the ribosomal protein S15Aa(e.g. an insect orthologue of the CG2033 Dm protein), said target genebeing represented by SEQ ID NOs. 150 and 151. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO. 339.

In one embodiment, the target gene encodes the ribosomal protein L19(e.g. an insect orthologue of the CG2746 Dm protein), said target genebeing represented by SEQ ID NOs. 200 and 201. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO.357.

In one embodiment, the target gene encodes the ribosomal protein L27(e.g. an insect orthologue of the CG4759 Dm protein), said target genebeing represented by SEQ ID NO. 386. In a preferred embodiment, theinsect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100%amino acid sequence identity to SEQ ID NO.390.

In one embodiment, the target gene encodes the mitochondrial cytochromec oxidase subunit II protein (e.g. an insect orthologue of the CG34069Dm protein), said target gene being represented by SEQ ID NO 25 and 26.In a preferred embodiment, the insect orthologue has at least 85%, 90%,92%, 94%, 96%, 98%, 99% or 100% amino acid sequence identity to SEQ IDNO. 91.

In one embodiment, the target gene encodes the ATP synthase-γ chain(e.g. an insect orthologue of the CG7610 Dm protein), said target genebeing represented by SEQ ID NOs 129 and 138. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO. 338.

In one embodiment, the target gene encodes the ubiquitin-5E (e.g. aninsect orthologue of the CG32744 Dm protein) said target gene beingrepresented by SEQ ID NOs. 186 and 187, 202 and 203. In a preferredembodiment, the insect orthologue has at least 85%, 90%, 92%, 94%, 96%,98%, 99% or 100% amino acid sequence identity to one or both of SEQ IDNOs.355 and 358.

In one embodiment, the target gene encodes the proteasome beta-typesubunit (e.g. an insect orthologue of the CG17331 Dm protein) saidtarget gene being represented by SEQ ID NO. 387. In a preferredembodiment, the insect orthologue has at least 85%, 90%, 92%, 94%, 96%,98%, 99% or 100% amino acid sequence identity to SEQ ID NO.391.

In one embodiment, the target gene encodes the protein which is aninsect orthologue of the CG13704 Dm protein, said target gene beingrepresented by SEQ ID NO.388. In a preferred embodiment, the insectorthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100% aminoacid sequence identity to SEQ ID NO.392.

In one embodiment, the target gene encodes the Rpn12 protein (e.g. aninsect orthologue of the CG4157 Dm protein) said target gene beingrepresented by SEQ ID NO. 389. In a preferred embodiment, the insectorthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100% aminoacid sequence identity to SEQ ID NO.393.

In accordance with a second aspect of the invention, there is provided acomposition for preventing and/or controlling insect pest infestationcomprising at least one interfering ribonucleic acid (RNA) and at leastone suitable carrier, excipient or diluent, wherein the interfering RNAfunctions upon uptake by the pest to down-regulate the expression of atarget gene within said pest,

wherein the RNA comprises at least one silencing element wherein thesilencing element is a region of double-stranded RNA comprising annealedcomplementary strands, one strand of which comprises or consists of asequence of nucleotides which is at least partially complementary to atarget nucleotide sequence within the target gene, and wherein thetarget gene(i) is selected from the group of genes having a nucleotide sequencecomprising any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13,14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62,19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78,143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135,226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153,246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389,or the complement thereof, or having a nucleotide sequence so that, whenthe two sequences are optimally aligned and compared, is at least 75%identical to any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189,27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6,35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50,13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132,214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126,135, 226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234to 237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152,153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159,258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167,166, 270 to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200,201, 314 to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387,388, 389, or the complement thereof, or(ii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, or having a nucleotide sequence so that, whensaid gene comprising said fragment is optimally aligned and comparedwith any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14,51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19,20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143,121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301,145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237,302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389,said nucleotide sequence is at least 75% identical to any of SEQ ID NOs1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204,16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21,22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142, 176,182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144,178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133,218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127,148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302 to 305,129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to 249, 154,155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161,262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273, 168,170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to 317, 186,202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, or thecomplement thereof, or(iii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, and wherein when said fragment is optimallyaligned and compared with the corresponding fragment in any of SEQ IDNOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15,204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66,21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142,176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122,144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124,133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229,127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302 to305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to 249,154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160,161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273,168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to 317,186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, saidnucleotide sequence of said fragment is at least 75% identical to saidcorresponding fragment of any of SEQ ID NOs 1, 174, 404, 180, 188, 2,175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11,12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325,17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74,25, 26, 75 to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to 209,286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128, 149,184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150, 151,242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to269, 165, 167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172, 173, 278to 281, 200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309, 318 to321, 386, 387, 388, 389, or the complement thereof, or(iv) is an insect pest orthologue of a gene having a nucleotide sequencecomprising any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13,14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62,19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78,143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135,226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153,246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389,or the complement thereof, wherein the two orthologous genes are similarin sequence to such a degree that when the two genes are optimallyaligned and compared, the orthologue has a sequence that is at least 75%identical to any of the sequences represented by SEQ ID NOs 1, 174, 404,180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70,23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142, 176, 182, 130, 177,183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179,210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146,125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to233, 128, 149, 184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238 to241, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253,156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163,162, 164, 266 to 269, 165, 167, 166, 270 to 273, 168, 170, 169, 274 to277, 172, 173, 278 to 281, 200, 201, 314 to 317, 186, 202, 187, 203, 306to 309, 318 to 321, 386, 387, 388, 389, or(v) is selected from the group of genes having a nucleotide sequenceencoding an amino acid sequence that, when the two amino acid sequencesare optimally aligned and compared, is at least 85% identical to theamino acid sequence encoded by any of SEQ ID NOs 1, 174, 404, 180, 188,2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141,11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to74, 25, 26, 75 to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128,149, 184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150,151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157,254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164,266 to 269, 165, 167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172,173, 278 to 281, 200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309,318 to 321, 386, 387, 388, 389.

The composition of the invention may be used for the prevention and/orcontrol of pest infestation. In certain embodiments, the composition maybe used as a pesticide for a plant or for propagation or reproductivematerial of a plant. In a further aspect, provided herein is acombination for preventing and/or controlling pest infestationcomprising the composition of the invention and at least one otheractive agent.

In a further aspect, provided herein is a method for down-regulatingexpression of a target gene in an insect pest species in order toprevent and/or control pest infestation, comprising contacting said pestspecies with an effective amount of at least one interfering ribonucleicacid (RNA), wherein the interfering RNA functions upon uptake by thepest to down-regulate the expression of a target gene within said pest,wherein the RNA comprises at least one silencing element wherein thesilencing element is a region of double-stranded RNA comprising annealedcomplementary strands, one strand of which comprises or consists of asequence of nucleotides which is at least partially complementary to atarget nucleotide sequence within the target gene, and wherein thetarget gene

(i) is selected from the group of genes having a nucleotide sequencecomprising any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13,14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62,19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78,143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135,226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153,246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389,or the complement thereof, or having a nucleotide sequence so that, whenthe two sequences are optimally aligned and compared, is at least 75%preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, or(ii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, or having a nucleotide sequence so that, whensaid gene comprising said fragment is optimally aligned and comparedwith any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14,51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19,20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143,121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301,145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237,302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389,said nucleotide sequence is at least 75% preferably at least 80%, 85%,90%, 95%, 98% or 99% identical to any of SEQ ID NOs 1, 174, 404, 180,188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3,4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46,141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71to 74, 25, 26, 75 to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128,149, 184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150,151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157,254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164,266 to 269, 165, 167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172,173, 278 to 281, 200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309,318 to 321, 386, 387, 388, 389, or the complement thereof, or(iii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, and wherein when said fragment is optimallyaligned and compared with the corresponding fragment in any of SEQ IDNOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15,204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66,21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142,176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122,144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124,133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229,127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302 to305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to 249,154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160,161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273,168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to 317,186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, saidnucleotide sequence of said fragment is at least 75% preferably at least80%, 85%, 90%, 95%, 98% or 99% identical to said corresponding fragmentof any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30,282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38,140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20,63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143,121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301,145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237,302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, or(iv) is an insect pest orthologue of a gene having a nucleotide sequencecomprising any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13,14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62,19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78,143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135,226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153,246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389,or the complement thereof, wherein the two orthologous genes are similarin sequence to such a degree that when the two genes are optimallyaligned and compared, the orthologue has a sequence that is at least 75%preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to any ofthe sequences represented by SEQ ID NOs 1, 174, 404, 180, 188, 2, 175,181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34,139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12,47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17,18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25,26, 75 to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293,123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225,147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184,137, 185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257,158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269,165, 167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to281, 200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309, 318 to 321,386, 387, 388, 389, or(v) is selected from the group of genes having a nucleotide sequenceencoding an amino acid sequence that, when the two amino acid sequencesare optimally aligned and compared, is at least 85% preferably at least90%, 95%, 98% or 99% identical to the amino acid sequence encoded by anyof SEQ ID NOs. SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14,51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19,20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143,121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301,145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237,302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389.

In accordance with a further aspect of the invention, there is providedan isolated polynucleotide selected from the group consisting of:

(i) a polynucleotide which comprises at least 21, preferably at least22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 70, 80, 90,100, 110, 125, 150, 175, 200, 225, 250, 300, 350, 400, 450, 500, 550,600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 2000 or 3000contiguous nucleotides of a nucleotide sequence as represented by any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, or(ii) a polynucleotide which consists of at least 21, preferably at least22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 70, 80, 90,100, 110, 125, 150, 175, 200, 225, 250, 300, 350, 400, 450, 500, 550,600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 2000 or 3000contiguous nucleotides of a nucleotide sequence as represented by any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, or(iii) a polynucleotide which comprises at least 21, preferably at least22, 23 or 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 70, 80,90, 100, 110, 125, 150, 175, 200, 225, 250, 300, 350, 400, 450, 500,550, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 2000 or3000 contiguous nucleotides of a nucleotide sequence as represented inany of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38,140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20,63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143,121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301,145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237,302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, that, when the two sequences are optimallyaligned and compared, said polynucleotide is at least 75% preferably atleast 80%, 85%, 90%, 95%, 98% or 99% identical to any of SEQ ID NOs 1,174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297,310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9,10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205,55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142, 176, 182, 130,177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131,179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221,146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230to 233, 128, 149, 184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238to 241, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265,163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273, 168, 170, 169, 274to 277, 172, 173, 278 to 281, 200, 201, 314 to 317, 186, 202, 187, 203,306 to 309, 318 to 321, 386, 387, 388, 389, or the complement thereof,or(iv) a polynucleotide which comprises a fragment of at least 21,preferably at least 22, 23 or 24, 25, 26, 27, 28, 29, 30, 35, 40, 45,50, 55, 60, 70, 80, 90, 100, 110, 125, 150, 175, 200, 225, 250, 300,350, 400, 450, 500, 550, 600, 700, 800, 900, 1000, 1100, 1200, 1300,1400, 1500, 2000 or 3000 contiguous nucleotides of a nucleotide asrepresented in any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181,189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139,5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289,298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123,132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147,126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137,185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245,152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158,159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165,167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281,200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386,387, 388, 389, or the complement thereof, and wherein said fragment orsaid complement has a nucleotide sequence that, when said fragment isoptimally aligned and compared with the corresponding fragment in any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389,said nucleotide sequence is at least 75% preferably at least 80%, 85%,90%, 95%, 98% or 99% identical to said corresponding fragment of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389 orthe complement thereof, or(v) a polynucleotide which consists of a fragment of at least 21,preferably at least 22, 23 or 24, 25, 26, 27, 28, 29, 30, 35, 40, 45,50, 55, 60, 70, 80, 90, 100, 110, 125, 150, 175, 200, 225, 250, 300,350, 400, 450, 500, 550, 600, 700, 800, 900, 1000, 1100, 1200, 1300,1400, 1500, 2000 or 3000 contiguous nucleotides of a nucleotide asrepresented in any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181,189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139,5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289,298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123,132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147,126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137,185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245,152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158,159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165,167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281,200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386,387, 388, 389, or the complement thereof, and wherein said fragment orsaid complement has a nucleotide sequence that, when said fragment isoptimally aligned and compared with the corresponding fragment in any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389,said nucleotide sequence is at least 75% preferably at least 80%, 85%,90%, 95%, 98% or 99% identical to said corresponding fragment of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389 orthe complement thereof, or(vi) a polynucleotide encoding an amino acid sequence that, when the twoamino acid sequences are optimally aligned and compared, is at least 70%preferably at least 75%, 80%, 85%, 90%, 95%, 98% or 99% identical to theamino acid sequence encoded by any of SEQ ID NOs 1, 174, 404, 180, 188,2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141,11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to74, 25, 26, 75 to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128,149, 184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150,151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157,254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164,266 to 269, 165, 167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172,173, 278 to 281, 200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309,318 to 321, 386, 387, 388, 389, and wherein said polynucleotide is nolonger than 10000, 9000, 8000, 7000, 6000, 5000, 4000, 3000, 2000 or1500 nucleotides.

The amino acid sequences encoded by the target genes of the presentinvention are represented by SEQ ID NOs 79, 349, 405, 352, 356, 80, 326,81, 327, 82, 83, 328, 84, 329, 85, 86, 359, 87 to 91, 330, 350, 353,331, 351, 332 to 336, 337, 354, 338 to 344, 346, 345, 347, 348, 357,355, 358, 390 to 393, respectively.

In a particular aspect of the invention, the isolated polynucleotide ispart of an interfering RNA molecule, typically part of the silencingelement, comprising at least one double-stranded region comprising asense RNA strand annealed by complementary basepairing to an antisenseRNA strand wherein the sense strand of the dsRNA molecule comprises asequence of nucleotides complementary to a sequence of nucleotideslocated within the RNA transcript of the target gene. More particularly,the isolated polynucleotide is cloned in a DNA construct in a sense andantisense orientation so that the upon transcription of the sense andantisense polynucleotide a dsRNA molecule is formed, which functionsupon uptake by a pest to inhibit or down-regulate the expression of atarget gene within said pest.

In one embodiment the present invention relates to an isolatedpolynucleotide that is cloned in a DNA construct in a sense andantisense orientation so that the upon transcription of the sense andantisense polynucleotide a dsRNA molecule is formed, which functionsupon uptake by an insect to inhibit or down-regulate the expression of atarget gene within the troponin/myofilament complex.

In one embodiment, the target gene encodes an insect wings up A(troponin I) protein (e.g. an insect orthologue of the CG7178 Dmprotein), said target gene being represented by SEQ ID NOs 1, 2, 174,404, 175, 180, 181, 188 and 189. In a preferred embodiment, the insectorthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100% aminoacid sequence identity to one or more of SEQ ID NOs. 79, 349, 405, 352or 356.

In one embodiment, the target gene encodes an upheld protein (e.g. aninsect orthologue of the CG7107 Dm protein), said target gene beingrepresented by SEQ ID NOs 121, 130, 142, 143, 176, 177, 182 and 183. Ina preferred embodiment, the insect orthologue has at least 85%, 90%,92%, 94%, 96%, 98%, 99% or 100% amino acid sequence identity to one ormore of SEQ ID NOs. 330, 350 or 353.

In one embodiment, the target gene encodes the tropomyosin 1 protein(e.g. an insect orthologue of the CG4898 Dm protein), or the tropomyosin2 protein (e.g. an insect orthologue of the CG4843 Dm protein), saidtarget gene being represented by SEQ ID NOs 123 and 132. In a preferredembodiment, the insect orthologue has at least 85%, 90%, 92%, 94%, 96%,98%, 99% or 100% amino acid sequence identity to SEQ ID NO. 332.

In one embodiment, the target gene encodes the myosin heavy chain (e.g.an insect orthologue of the CG17927 Dm protein), said target gene beingrepresented by SEQ ID NOs 122, 131, 144, 145, 178 and 179. In apreferred embodiment, the insect orthologue has at least 85%, 90%, 92%,94%, 96%, 98%, 99% or 100% amino acid sequence identity to one or moreof SEQ ID NOs. 331 or 351.

In one embodiment, the target gene encodes the myosin light chaincytoplasmic protein (e.g. an insect orthologue of the CG3201 Dmprotein), said target gene being represented by SEQ ID NOs 124 and 133.In a preferred embodiment, the insect orthologue has at least 85%, 90%,92%, 94%, 96%, 98%, 99% or 100% amino acid sequence identity to SEQ IDNO. 333.

In one embodiment, the target gene encodes the spaghetti squash protein(e.g. an insect orthologue of the CG3595 Dm protein), said target genebeing represented by SEQ ID NOs 125 and 134. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% identity to SEQ ID NO. 334.

In one embodiment, the target gene encodes the zipper protein (e.g. aninsect orthologue of the CG15792 Dm protein), said target gene beingrepresented by SEQ ID NOs 126 and 135. In a preferred embodiment, theinsect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100%identity to SEQ ID NO. 335.

In one embodiment, the target gene encodes the troponin C (e.g. aninsect orthologue of the CG2981, CG7930, CG9073, CG6514, CG12408,CG9073, CG7930, CG2981, CG12408 or CG6514 Dm protein), said target genebeing represented by SEQ ID NOs 127 and 136, or 128 and 137, or 184 and185. In a preferred embodiment, the insect orthologue has at least 85%,90%, 92%, 94%, 96%, 98%, 99% or 100% amino acid sequence identity to oneor more of SEQ ID NOs. 336, 337 and 354.

According to other embodiments, the present invention relates to anisolated polynucleotide that is cloned in a DNA construct in a sense andantisense orientation so that the upon transcription of the sense andantisense polynucleotide a dsRNA molecule is formed, which functionsupon uptake by an insect to inhibit or down-regulate the expression of atarget gene that encodes an insect ribosomal protein.

In one embodiment, the target gene encodes ribosomal protein S3A (e.g.an insect orthologue of the CG2168 Dm protein), said target gene beingrepresented by SEQ ID NOs 11, 12 and 141. In a preferred embodiment, theinsect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100%amino acid sequence identity to one or both of SEQ ID NO. 84 or 328.

In one embodiment, the target gene encodes the ribosomal protein LP1(e.g. an insect orthologue of the CG4087 Dm protein), said target genebeing represented by SEQ ID NO 3 and 4. In a preferred embodiment, theinsect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100%amino acid sequence identity to SEQ ID NO.80.

In one embodiment, the target gene encodes the ribosomal protein S3(e.g. an insect orthologue of the CG6779 Dm protein), said target genebeing represented by SEQ ID NOs 7 and 8. In a preferred embodiment, theinsect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100%amino acid sequence identity to SEQ ID NO.82.

In one embodiment, the target gene encodes the ribosomal protein L10Ab(e.g. an insect orthologue of the CG7283 Dm protein) represented by SEQID NOs 9 and 10. In a preferred embodiment, the insect orthologue has atleast 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100% amino acid sequenceidentity to SEQ ID NO. 83.

In one embodiment, the target gene encodes the ribosomal protein S18(e.g. an insect orthologue of the CG8900 Dm protein), said target genebeing represented by SEQ ID NO 13 and 14. In a preferred embodiment, theinsect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100%amino acid sequence identity to SEQ ID NO.85.

In one embodiment, the target gene encodes the ribosomal protein L4(e.g. an insect orthologue of the CG5502 Dm protein), said target generepresented by SEQ ID NO 5 and 6. In a preferred embodiment, the insectorthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100% aminoacid sequence identity to SEQ ID NO.81.

In one embodiment, the target gene encodes the ribosomal protein S27(e.g. an insect orthologue of the CG10423 Dm protein), said target genebeing represented by SEQ ID NO 15 and 16, 204 and 205. In a preferredembodiment, the insect orthologue has at least 85%, 90%, 92%, 94%, 96%,98%, 99% or 100% amino acid sequence identity to one or both of SEQ IDNOs.86 and 359.

In one embodiment, the target gene encodes the ribosomal protein L6(e.g. an insect orthologue of the CG11522 Dm protein), said target genebeing represented by SEQ ID NO 17 and 18. In a preferred embodiment, theinsect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100%amino acid sequence identity to SEQ ID NO. 87.

In one embodiment, the target gene encodes the ribosomal protein S13(e.g. an insect orthologue of the CG13389 Dm protein), said target genebeing represented by SEQ ID NO 19 and 20. In a preferred embodiment, theinsect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100%amino acid sequence identity to SEQ ID NO. 88.

In one embodiment, the target gene encodes the ribosomal protein L12(e.g. an insect orthologue of the CG3195 Dm protein), said target genebeing represented by SEQ ID NOs 21 and 22. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO.89.

In one embodiment, the target gene encodes the ribosomal protein L26(e.g. an insect orthologue of the CG6846 Dm protein), said target genebeing represented by SEQ ID NOs 158 and 159. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO. 343.

In one embodiment, the target gene encodes the ribosomal protein L21(e.g. an insect orthologue of the CG12775 Dm protein), said target genebeing represented by SEQ ID NO 165, 166 and 167. In a preferredembodiment, the insect orthologue has at least 85%, 90%, 92%, 94%, 96%,98%, 99% or 100% amino acid sequence identity to SEQ ID NOs 347 and 348.

In one embodiment, the target gene encodes the ribosomal protein S12(e.g. an insect orthologue of the CG11271 Dm protein), said target genebeing represented by SEQ ID NOs 156 and 157. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO. 342.

In one embodiment, the target gene encodes the ribosomal protein S28b(e.g. an insect orthologue of the CG2998 Dm protein), said target genebeing represented by SEQ ID NOs 160 and 161. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO. 344.

In one embodiment, the target gene encodes the ribosomal protein L13(e.g. an insect orthologue of the CG4651 Dm protein), said target genebeing represented by SEQ ID NOs. 154 and 155. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO. 341.

In one embodiment, the target gene encodes the ribosomal protein L10(e.g. an insect orthologue of the CG17521 Dm protein), said target genebeing represented by SEQ ID NOs. 163 and 164. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO. 345.

In one embodiment, the target gene encodes the ribosomal protein L5(e.g. an insect orthologue of the CG17489 Dm protein), said target genebeing represented by SEQ ID NOs. 152 and 153. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO. 340.

In one embodiment, the target gene encodes the ribosomal protein S15Aa(e.g. an insect orthologue of the CG2033 Dm protein), said target genebeing represented by SEQ ID NOs. 150 and 151. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO. 339.

In one embodiment, the target gene encodes the ribosomal protein L19(e.g. an insect orthologue of the CG2746 Dm protein), said target genebeing represented by SEQ ID NOs. 200 and 201. In a preferred embodiment,the insect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or100% amino acid sequence identity to SEQ ID NO.357.

In one embodiment, the target gene encodes the ribosomal protein L27(e.g. an insect orthologue of the CG4759 Dm protein), said target genebeing represented by SEQ ID NO. 386. In a preferred embodiment, theinsect orthologue has at least 85%, 90%, 92%, 94%, 96%, 98%, 99% or 100%amino acid sequence identity to SEQ ID NO.390.

Preferably, the methods of the invention find practical application inthe prevention and/or control of insect pest infestation, in particular,control of pest infestation of crop plants such as but not limited tocotton, potato, rice, strawberries, alfalfa, soy, tomato, canola,sunflower, sorghum, pearl millet, corn, eggplant, pepper and tobacco. Inaddition, the interfering RNA of the invention may be introduced intothe plants to be protected by routine genetic engineering techniques.

In all aspects of the invention, in preferred embodiments the targetgene

(i) is selected from the group of genes having a nucleotide sequencecomprising any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27to 30, 282 to 285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177,183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179,210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146,125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to233, or the complement thereof, or having a nucleotide sequence so that,when the two sequences are optimally aligned and compared, is at least75% preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to anyof SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, or thecomplement thereof, or(ii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, or thecomplement thereof, or having a nucleotide sequence so that, when saidgene comprising said fragment is optimally aligned and compared with anyof SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, saidnucleotide sequence is at least 75% preferably at least 80%, 85%, 90%,95%, 98% or 99% identical to any of SEQ ID NOs 1, 174, 404, 180, 188, 2,175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 121, 142,176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122,144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124,133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229,127, 148, 136, 230 to 233, or the complement thereof, or(iii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233 or thecomplement thereof, and wherein when said fragment is optimally alignedand compared with the corresponding fragment in any of SEQ ID NOs 1,174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297,310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289,298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123,132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147,126, 135, 226 to 229, 127, 148, 136, 230 to 233 said nucleotide sequenceof said fragment is at least 75% preferably at least 80%, 85%, 90%, 95%,98% or 99% identical to said corresponding fragment of any of SEQ ID NOs1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to297, 310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293,123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225,147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233 or the complementthereof, or(iv) is an insect pest orthologue of a gene having a nucleotide sequencecomprising any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27to 30, 282 to 285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177,183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179,210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146,125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to233, or the complement thereof, wherein the two orthologous genes aresimilar in sequence to such a degree that when the two genes areoptimally aligned and compared, the orthologue has a sequence that is atleast 75% preferably at least 80%, 85%, 90%, 95%, 98% or 99% identicalto any of the sequences represented by SEQ ID NOs 1, 174, 404, 180, 188,2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, or(v) is selected from the group of genes having a nucleotide sequenceencoding an amino acid sequence that, when the two amino acid sequencesare optimally aligned and compared, is at least 85% preferably at least90%, 95%, 98% or 99% identical to the amino acid sequence encoded by anyof SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233.

These target genes encode proteins within the troponin/myofilamentcomplex.

In all aspects of the invention, in preferred embodiments, the targetgene

(i) is selected from the group of genes having a nucleotide sequencecomprising any of SEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 150, 151, 242 to 245, 152, 153, 246 to 249,154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160,161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273,or the complement thereof, or having a nucleotide sequence so that, whenthe two sequences are optimally aligned and compared, is at least 75%preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to any ofSEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9,10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205,55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to70, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253,156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163,162, 164, 266 to 269, 165, 167, 166, 270 to 273, or the complementthereof, or(ii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9,10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205,55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to70, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253,156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163,162, 164, 266 to 269, 165, 167, 166, 270 to 273, or the complementthereof, or having a nucleotide sequence so that, when said genecomprising said fragment is optimally aligned and compared with any ofSEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9,10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205,55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to70, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253,156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163,162, 164, 266 to 269, 165, 167, 166, 270 to 273, said nucleotidesequence is at least 75% preferably at least 80%, 85%, 90%, 95%, 98% or99% identical to any of SEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38,140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20,63 to 66, 21, 22, 67 to 70, 150, 151, 242 to 245, 152, 153, 246 to 249,154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160,161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273,or the complement thereof, or(iii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9,10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205,55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to70, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253,156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163,162, 164, 266 to 269, 165, 167, 166, 270 to 273, or the complementthereof, and wherein when said fragment is optimally aligned andcompared with the corresponding fragment in any of SEQ ID NOs. 3, 4, 31to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141,11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 150, 151, 242to 245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to269, 165, 167, 166, 270 to 273, said nucleotide sequence of saidfragment is at least 75% preferably at least 80%, 85%, 90%, 95%, 98% or99% identical to said corresponding fragment of any of SEQ ID NOs. 3, 4,31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46,141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 150, 151,242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to269, 165, 167, 166, 270 to 273, or the complement thereof, or(iv) is an insect pest orthologue of a gene having a nucleotide sequencecomprising any of SEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 150, 151, 242 to 245, 152, 153, 246 to 249,154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160,161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273,or the complement thereof, wherein the two orthologous genes are similarin sequence to such a degree that when the two genes are optimallyaligned and compared, the orthologue has a sequence that is at least 75%preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to any ofthe sequences represented by SEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13,14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62,19, 20, 63 to 66, 21, 22, 67 to 70, 150, 151, 242 to 245, 152, 153, 246to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270to 273, or(v) is selected from the group of genes having a nucleotide sequenceencoding an amino acid sequence that, when the two amino acid sequencesare optimally aligned and compared, is at least 85% identical to theamino acid sequence encoded by any of SEQ ID NOs. 3, 4, 31 to 34, 139,5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 150, 151, 242 to 245, 152,153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159,258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167,166, 270 to 273.

These target genes encode insect ribosomal proteins.

In all aspects of the invention, in preferred embodiments, the targetgene (i) is selected from the group of genes having a nucleotidesequence comprising any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175,181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, or thecomplement thereof, or having a nucleotide sequence so that, when thetwo sequences are optimally aligned and compared, is at least 75%preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, or the complement thereof, or

(ii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, or the complement thereof, or having anucleotide sequence so that, when said gene comprising said fragment isoptimally aligned and compared with any of SEQ ID NOs 1, 174, 404, 180,188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313,said nucleotide sequence is at least 75% preferably at least 80%, 85%,90%, 95%, 98% or 99% identical to any of SEQ ID NOs 1, 174, 404, 180,188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, orthe complement thereof, or(iii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, or the complement thereof, and wherein whensaid fragment is optimally aligned and compared with the correspondingfragment in any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189,27 to 30, 282 to 285, 294 to 297, 310 to 313, said nucleotide sequenceof said fragment is at least 75% preferably at least 80%, 85%, 90%, 95%,98% or 99% identical to said corresponding fragment of any of SEQ ID NOs1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to297, 310 to 313, or the complement thereof, or(iv) is an insect pest orthologue of a gene having a nucleotide sequencecomprising any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27to 30, 282 to 285, 294 to 297, 310 to 313, or the complement thereof,wherein the two orthologous genes are similar in sequence to such adegree that when the two genes are optimally aligned and compared, theorthologue has a sequence that is at least 75% preferably at least 80%,85%, 90%, 95%, 98% or 99% identical to any of the sequences representedby SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, or(v) is selected from the group of genes having a nucleotide sequenceencoding an amino acid sequence that, when the two amino acid sequencesare optimally aligned and compared, is at least 85% preferably at least90%, 95%, 98% or 99% identical to the amino acid sequence encoded by anyof SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313.

In preferred embodiments, this target gene may encode an insect thetroponin I protein (e.g. an insect orthologue of the CG7178 Dm protein).The insect troponin I protein may have an amino acid sequence which isat least 85%, 90%, 92%, 94%, 96%, 98%, 99% identical to the amino acidsequence as presented in any of SEQ ID NOs 79, 349, 405, 352 or 356(when said encoded proteins are optimally aligned).

BRIEF DESCRIPTION OF THE TABLES AND FIGURES

Table 1 Lygus hesperus novel targets identified from first screen.

Table 1B Lygus hesperus novel targets in Lh594 pathway.

Table 1C Lygus hesperus novel targets identified from second roundscreen.

Table 2 Polynucleotide sequences of target genes identified in Lygushesperus.

Table 3 Amino acid sequences of target genes identified in Lygushesperus.

Table 4 dsRNAs (sense strand represented by equivalent DNA sequence)corresponding to Lygus hesperus target genes and primers for producingthe dsRNAs.

Table 5 Lygus hesperus targets ranking according to dose response curves(DRCs) and compared to bench mark targets Lh423 & Lh105.

Table 6 Lygus hesperus targets from second round screen-rankingaccording to DRCs and compared to bench mark targets Lh423 & Lh594.

Table 7 Polynucleotide sequences of target genes identified in Coloradopotato beetle (CPB).

Table 8 Amino acid sequences of target genes identified in CPB.

Table 9 dsRNAs (sense strand represented by equivalent DNA sequence)corresponding to CPB target genes and primers for producing the dsRNAs.

Table 10 Polynucleotide sequences of target genes identified in brownplant hopper (BPH).

Table 11 Amino acid sequences of target genes identified in BPH.

Table 12 dsRNAs (sense strand represented by equivalent DNA sequence)corresponding to BPH target genes and primers for producing the dsRNAs.

Table 13 Primers used for amplification of aphid cDNAs, based on peaaphid genomic sequence.

Table 14 Polynucleotide sequences of target genes identified in aphids.

Table 15 Amino acid sequences of target genes identified in aphids.

Table 16 dsRNAs (sense strand represented by equivalent DNA sequence)corresponding to aphid target genes and primers for producing thedsRNAs.

Table 17 Degenerate primers used for amplification of CPB Ld594 cDNA

Table 18 Degenerate primers used for amplification of BPH cDNAs

Table 19: Leptinotarsa decemlineata novel targets from the screen.

Table 20: Nilaparvata lugens novel identified target.

Table 21: Acyrthosiphon pisum novel identified targets.

FIG. 1: Plates Lh001_009 second confirmation assay. Dark bars: mortalityat day 3 to 6, light bars: mortality at day 6 to 8. Candidate clones arenamed using the “Lygxxx” screening codes and the “Lhxxx” targetnomenclature codes.

FIG. 2: Plates Lh010_020 second confirmation assay. Dark bars: mortalityat day 3 to 6, light bars: mortality at day 6 to 8. Candidate clones arenamed using the “Lygxxx” screening codes and the “Lhxxx” targetnomenclature codes.

FIG. 3: Mortality analysis of Lygus novel targets from plates Lh001 toLh009, expressed as % mortality over a 10 day period. Controls areindicated in dotted lines. Positive control: Lh423 dsRNA (RpL19).Negative controls: GFP dsRNA and diet only (Control).

FIG. 4: Mortality analysis of Lygus novel targets from plates Lh010 toLh020, expressed as % mortality over a 10 day period. Controls areindicated in dotted lines. Positive control: Lh423 (RpL19). Negativecontrols: GFP and diet only (Control).

FIGS. 5 to 9 Lygus hesperus novel targets—dose response curves atconcentrations of purified synthetic dsRNA ranging from 0.4 to 0.025μg/μl (in the figure, the unit “μg/μl” is not displayed). GFP dsRNA andmilliQ water were used negative controls. dsRNA of targets were producedusing the primers as described in the example section 1.1.

FIG. 10 Lh594 dose response curve, at dsRNA concentrations ranging from0.05 to 0.001 μg/μl. GFP dsRNA and milliQ water were used negativecontrols.

FIG. 11 A dsRNA activity in Lygus hesperus bioassay in absence of tRNA.Lh594 (5 μg/μl); positive control: Lh423 (5 μg/μl); negative controls:GFP dsRNA (5 μg/μl) and milliQ water; B Identification of Lh594 limit ofactivity using decreasing concentration of dsRNA (from 5 μg to 0.25 μg).Negative controls: GFP dsRNA (5 μg/μl) and milliQ water.

FIG. 12 Plates Lh010 to Lh020 second confirmation assay of second screentargets. Dark bars: mortality at day 4 to 8, light bars: mortality atday 4 to 6. Candidate clones are named using the “Lygxxx” screeningcodes and the “Lhxxx” target nomenclature codes.

FIG. 13 Assay results for Lygus troponin pathway targets, tested at 0.5μg/μl fixed.

FIGS. 14 A-B Lygus hesperus novel targets from troponin pathway—doseresponse curves at concentrations of purified synthetic dsRNA rangingfrom 0.4 to 0.025 μg/μl (in the figure, the unit “μg/μl” is not alwaysdisplayed). GFP dsRNA and milliQ water were used as negative controls.

FIGS. 15 A-D Lygus hesperus novel targets of second screen targets—doseresponse curves at concentrations of purified synthetic dsRNA rangingfrom 0.5 to 0.05 μg/μl. GFP dsRNA and milliQ water were used as negativecontrols.

FIG. 16 Survival analysis of CPB larvae treated with 1 μg dsRNA Ld594,Ld619 and Ld620. Positive controls included 1 μg dsRNA of bench marktargets Ld513 and Ld049. Negative controls included milliQ water and FP.

FIG. 17 Effects of Ld594, Ld619 and Ld620 dsRNAs on pupation of CPB4^(th) instar larvae, compared to untreated control (UTC). Bugs were fed1 μg dsRNA dispensed in potato leaf disks, then were allowed to feed onuntreated potato leaves (A) for 4 days before being placed onvermiculite. To assess the effect of the dsRNA, dead insects wereexcavated from the vermiculite (because of the strong effects induced byLd594 dsRNA, no pupae could be recovered from the vermiculite andtherefore, no image is available for this target dsRNA) (B).

FIG. 18 Effect of CPB Ld594, 619 & 620 dsRNAs on survival and fitness ofCPB adults. Assessments were performed on days 4, 6, 7, 8, 11 and 13.Control MQ: milliQ water.

FIG. 19 Activity of dsRNA from NI594 pathway in brown plant hopper.DsRNAs were tested at 0.5 μg/μl in presence of 0.1% CHAPSO. Positivecontrol: NI537 dsRNA (0.5 μg/μl), negative controls: GFP dsRNA (0.5μg/μl) and diet alone.

FIG. 20 Activity of dsRNA from Ap594, Ap423, Ap537 and Ap560 on A.pisum. DsRNAs were tested at 0.5 μg/μl in presence of 5 μg/μl tRNA.Negative control: GFP dsRNA (0.5 μg/μl).

FIG. 21 Mortality percentages of L. decemlineata larvae on artificialdiet treated with dsRNA. Ld583, Ld584, Ld586 & Ld588 represent targetclones. Positive control: Ld513; negative control: FP.

DETAILED DESCRIPTION OF THE INVENTION

The present inventors have discovered that down-regulating theexpression of particular target genes in insect pest species by RNAi canbe used to effectively prevent and/or control infestation by said insectpest.

As used herein, the term “control” of pest infestation refers to anyeffect on a pest that serves to limit and/or reduce either the numbersof pest organisms and/or the damage caused by the pest.

Preferred target genes are therefore essential genes that control orregulate one or more essential biological functions within the insectpest, for example, cell division, reproduction, energy metabolism,digestion, neurological function and the like. Down-regulation of theseessential genes by RNAi techniques can lead to death of the insect, orotherwise significantly retard growth and development or impair theability of the pest to colonize an environment or infest host organisms.

The present inventors have now identified superior target genes ofinsect pest species belonging to the Lygus, Leptinotarsa, Nilaparvataand Acyrthosiphum genus, which targets are envisaged for use singly orin combination as an effective means for RNAi-mediated control of insectinfestation, for example of agronomically important crops. Orthologuesof these newly identified target genes can be used in other insectspecies to control pest infestation of the corresponding relevant crops.

More specifically, the present inventors describe here that genesencoding for proteins of the troponin/myofilament complex form excellenttarget genes for suppression by the RNA inhibition machinery. One ofthese target genes encoded the insect troponin I protein (wings up A)which is an orthologue of the Drosophila CG7178 protein. This protein isinvolved in muscular contraction and belongs to a physiological pathwaythat was not yet fully explored for (insect) pest control through RNAinhibition. Moreover, since this protein complex is animal specific, noplant genes homologues or orthologues are known, reducing the risk ofoff-type plant phenotypes when expressing target dsRNA in plants. Inaddition, in Drosophila, troponin I is described as a haplo-insufficientgene, displaying a mutant phenotype in the heterozygote state. Suchgenes are particularly susceptible to reduced mRNA expression levels andas such can be considered as ideal RNAi targets.

Further interesting target genes in this troponin/myofilament complexare listed below.

Annotation ID Cytology Dm identifier up upheld CG7107 Tm1 tropomyosin 1CG4898 Tm2 tropomyosin 2 CG4843 Mhc myosin heavy chain CG17927 Mlc-cmyosin light chain cytoplasmic CG3201 sqh spaghetti squash CG3595 zipzipper CG15792

Thus, according to one embodiment the present invention relates to aninterfering ribonucleic acid (RNA) that functions upon uptake by aninsect pest species to down-regulate expression of a target gene in saidinsect pest, wherein the RNA comprises at least one silencing elementwherein the silencing element is a region of double-stranded RNAcomprising annealed complementary strands, one strand of which comprisesor consists of a sequence of nucleotides which is at least partiallycomplementary to a target nucleotide sequence within the target gene,and wherein the target gene

(i) is selected from the group of genes having a nucleotide sequencecomprising any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27to 30, 282 to 285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177,183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179,210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146,125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to233 or the complement thereof, or having a nucleotide sequence so that,when the two sequences are optimally aligned and compared, is at least75% preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to anyof SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, or thecomplement thereof, or(ii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, or thecomplement thereof, or having a nucleotide sequence so that, when saidgene comprising said fragment is optimally aligned and compared with anyof SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, saidnucleotide sequence is at least 75% preferably at least 80%, 85%, 90%,95%, 98% or 99% identical to any of SEQ ID NOs 1, 174, 404, 180, 188, 2,175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 121, 142,176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122,144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124,133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229,127, 148, 136, 230 to 233, or the complement thereof, or(iii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, or thecomplement thereof, and wherein when said fragment is optimally alignedand compared with the corresponding fragment in any of SEQ ID NOs 1,174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297,310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289,298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123,132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147,126, 135, 226 to 229, 127, 148, 136, 230 to 233, said nucleotidesequence of said fragment is at least 75% preferably at least 80%, 85%,90%, 95%, 98% or 99% identical to said corresponding fragment of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, or thecomplement thereof, or(iv) is an insect pest orthologue of a gene having a nucleotide sequencecomprising any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27to 30, 282 to 285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177,183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179,210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146,125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to233, or the complement thereof, wherein the two orthologous genes aresimilar in sequence to such a degree that when the two genes areoptimally aligned and compared, the orthologue has a sequence that is atleast 75% preferably at least 80%, 85%, 90%, 95%, 98% or 99% identicalto any of the sequences represented by SEQ ID NOs 1, 174, 404, 180, 188,2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, or(v) is selected from the group of genes having a nucleotide sequenceencoding an amino acid sequence that, when the two amino acid sequencesare optimally aligned and compared, is at least 85% preferably at least90%, 95%, 98% or 99% identical to the amino acid sequence encoded by anyof SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233.

In a preferred embodiment, the target gene encodes an insect proteinchosen from the troponin/myofilament complex chosen from the groupcomprising the troponin I (e.g. an insect orthologue of the CG7178 Dmprotein), the upheld protein (e.g. an insect orthologue of the CG7107 Dmprotein), the tropomyosin 1 protein (e.g. an insect orthologue of theCG4898 Dm protein), the tropomyosin 2 protein (e.g. an insect orthologueof the CG4843 Dm protein), the myosin heavy chain (e.g. an insectorthologue of the CG17927 Dm protein), the myosin light chaincytoplasmic protein (e.g. an insect orthologue of the CG3201 Dmprotein), the spaghetti squash protein (e.g. an insect orthologue of theCG3595 Dm protein), the zipper protein (e.g. an insect orthologue of theCG15792 Dm protein), the troponin C (e.g. an insect orthologue of theCG2981, CG7930, CG9073, CG6514, CG12408, CG9073, CG7930, CG2981, CG12408or CG6514 Dm protein)

In other embodiments, the present invention relates to an interferingribonucleic acid (RNA) that functions upon uptake by an insect pestspecies to down-regulate expression of a target gene in said insectpest, wherein the RNA comprises at least one silencing element whereinthe silencing element is a region of double-stranded RNA comprisingannealed complementary strands, one strand of which comprises orconsists of a sequence of nucleotides which is at least partiallycomplementary to a target nucleotide sequence within the target gene,and wherein the target gene

(i) is selected from the group of genes having a nucleotide sequencecomprising any of SEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 150, 151, 242 to 245, 152, 153, 246 to 249,154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160,161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273,or the complement thereof, or having a nucleotide sequence so that, whenthe two sequences are optimally aligned and compared, is at least 75%preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to any ofSEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9,10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205,55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to70, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253,156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163,162, 164, 266 to 269, 165, 167, 166, 270 to 273, or the complementthereof, or(ii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9,10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205,55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to70, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253,156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163,162, 164, 266 to 269, 165, 167, 166, 270 to 273, or the complementthereof, or having a nucleotide sequence so that, when said genecomprising said fragment is optimally aligned and compared with any ofSEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9,10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205,55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to70, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253,156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163,162, 164, 266 to 269, 165, 167, 166, 270 to 273, said nucleotidesequence is at least 75% preferably at least 80%, 85%, 90%, 95%, 98% or99% identical to any of SEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38,140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20,63 to 66, 21, 22, 67 to 70, 150, 151, 242 to 245, 152, 153, 246 to 249,154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160,161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273,or the complement thereof, or(iii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9,10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205,55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to70, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253,156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163,162, 164, 266 to 269, 165, 167, 166, 270 to 273, or the complementthereof, and wherein when said fragment is optimally aligned andcompared with the corresponding fragment in any of SEQ ID NOs. 3, 4, 31to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141,11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 150, 151, 242to 245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to269, 165, 167, 166, 270 to 273, said nucleotide sequence of saidfragment is at least 75% preferably at least 80%, 85%, 90%, 95%, 98% or99% identical to said corresponding fragment of any of SEQ ID NOs. 3, 4,31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46,141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 150, 151,242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to269, 165, 167, 166, 270 to 273, or the complement thereof, or(iv) is an insect pest orthologue of a gene having a nucleotide sequencecomprising any of SEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 150, 151, 242 to 245, 152, 153, 246 to 249,154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160,161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273,or the complement thereof, wherein the two orthologous genes are similarin sequence to such a degree that when the two genes are optimallyaligned and compared, the orthologue has a sequence that is at least 75%preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to any ofthe sequences represented by SEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13,14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62,19, 20, 63 to 66, 21, 22, 67 to 70, 150, 151, 242 to 245, 152, 153, 246to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270to 273, or(v) is selected from the group of genes having a nucleotide sequenceencoding an amino acid sequence that, when the two amino acid sequencesare optimally aligned and compared, is at least 85% preferably at least90%, 95%, 98% or 99% identical to the amino acid sequence encoded by anyof SEQ ID NOs. 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42,9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16,205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22,67 to 70, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265,163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273.

In a preferred embodiment, the target gene encodes an insect ribosomalprotein chosen from the group comprising the ribosomal protein S3A (e.g.an insect orthologue of the CG2168 Dm protein), the ribosomal proteinLP1 (e.g. an insect orthologue of the CG4087 Dm protein), the ribosomalprotein S3 (e.g. an insect orthologue of the CG6779 Dm protein), theribosomal protein L10Ab (e.g. an insect orthologue of the CG7283 Dmprotein), the ribosomal protein S18 (e.g. an insect orthologue of theCG8900 Dm protein), the ribosomal protein L4 (e.g. an insect orthologueof the CG5502 Dm protein), the ribosomal protein S27 (e.g. an insectorthologue of the CG10423 Dm protein), the ribosomal protein L6 (e.g. aninsect orthologue of the CG11522 Dm protein), the ribosomal protein S13(e.g. an insect orthologue of the CG13389 Dm protein), and the ribosomalprotein L12 (e.g. an insect orthologue of the CG3195 Dm protein), theribosomal protein L26 (e.g. an insect orthologue of the CG6846 Dmprotein), the ribosomal protein L21 (e.g. an insect orthologue of theCG12775 Dm protein), the ribosomal protein S12 (e.g. an insectorthologue of the CG11271 Dm protein), the ribosomal protein S28b (e.g.an insect orthologue of the CG2998 Dm protein), the ribosomal proteinL13 (e.g. an insect orthologue of the CG4651 Dm protein), the ribosomalprotein L10 (e.g. an insect orthologue of the CG17521 Dm protein), theribosomal protein L5 (e.g. an insect orthologue of the CG17489 Dmprotein), the ribosomal protein S15Aa (e.g. an insect orthologue of theCG2033 Dm protein), the ribosomal protein L19 (e.g. an insect orthologueof the CG2746 Dm protein), the ribosomal protein L27 (e.g. an insectorthologue of the CG4759 Dm protein)

In one embodiment, the present invention relates to an interferingribonucleic acid (RNA) that functions upon uptake by an insect pestspecies to down-regulate expression of a target gene in said insectpest, wherein the RNA comprises at least one silencing element whereinthe silencing element is a region of double-stranded RNA comprisingannealed complementary strands, one strand of which comprises orconsists of a sequence of nucleotides which is at least partiallycomplementary to a target nucleotide sequence within the target gene,and wherein the target gene

(i) is selected from the group of genes having a nucleotide sequencecomprising any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27to 30, 282 to 285, 294 to 297, 310 to 313, or the complement thereof, orhaving a nucleotide sequence so that, when the two sequences areoptimally aligned and compared, is at least 75% preferably at least 80%,85%, 90%, 95%, 98% or 99% identical to any of SEQ ID NOs 1, 174, 404,180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to313, or the complement thereof, or(ii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, or the complement thereof, or having anucleotide sequence so that, when said gene comprising said fragment isoptimally aligned and compared with any of SEQ ID NOs 1, 174, 404, 180,188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313,said nucleotide sequence is at least 75% preferably at least 80%, 85%,90%, 95%, 98% or 99% identical to any of SEQ ID NOs 1, 174, 404, 180,188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, orthe complement thereof, or(iii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, or the complement thereof, and wherein whensaid fragment is optimally aligned and compared with the correspondingfragment in any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189,27 to 30, 282 to 285, 294 to 297, 310 to 313, said nucleotide sequenceof said fragment is at least 75% preferably at least 80%, 85%, 90%, 95%,98% or 99% identical to said corresponding fragment of any of SEQ ID NOs1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to297, 310 to 313, or the complement thereof, or(iv) is an insect pest orthologue of a gene having a nucleotide sequencecomprising any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27to 30, 282 to 285, 294 to 297, 310 to 313, or the complement thereof,wherein the two orthologous genes are similar in sequence to such adegree that when the two genes are optimally aligned and compared, theorthologue has a sequence that is at least 75% preferably at least 80%,85%, 90%, 95%, 98% or 99% identical to any of the sequences representedby SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, or(v) is selected from the group of genes having a nucleotide sequenceencoding an amino acid sequence that, when the two amino acid sequencesare optimally aligned and compared, is at least 85% preferably at least90%, 95%, 98% or 99% identical to the amino acid sequence encoded by anyof SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313.

In one embodiment, the present invention relates to an interferingribonucleic acid (RNA) that functions upon uptake by an insect pestspecies to down-regulate expression of a target gene in said insectpest, wherein the RNA comprises at least one silencing element whereinthe silencing element is a region of double-stranded RNA comprisingannealed complementary strands, one strand of which comprises orconsists of a sequence of nucleotides which is at least partiallycomplementary to a target nucleotide sequence within the target gene,and wherein the target gene

(i) is selected from the group of genes having a nucleotide sequencecomprising any of SEQ ID NOs 141, 11, 12, or the complement thereof, orhaving a nucleotide sequence so that, when the two sequences areoptimally aligned and compared, is at least 75% preferably at least 80%,85%, 90%, 95%, 98% or 99% identical to any of SEQ ID NOs 141, 11, 12, orthe complement thereof, or(ii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 141, 11, 12, or the complement thereof, or having anucleotide sequence so that, when said gene comprising said fragment isoptimally aligned and compared with any of SEQ ID NOs 141, 11, 12, saidnucleotide sequence is at least 75% preferably at least 80%, 85%, 90%,95%, 98% or 99% identical to any of SEQ ID NOs 141, 11, 12, or thecomplement thereof, or(iii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 141, 11, 12, or the complement thereof, and wherein when saidfragment is optimally aligned and compared with the correspondingfragment in any of SEQ ID NOs 141, 11, 12, said nucleotide sequence ofsaid fragment is at least 75% preferably at least 80%, 85%, 90%, 95%,98% or 99% identical to said corresponding fragment of any of SEQ ID NOs141, 11, 12, or the complement thereof, or(iv) is an insect pest orthologue of a gene having a nucleotide sequencecomprising any of SEQ ID NOs 141, 11, 12, or the complement thereof,wherein the two orthologous genes are similar in sequence to such adegree that when the two genes are optimally aligned and compared, theorthologue has a sequence that is at least 75% preferably at least 80%,85%, 90%, 95%, 98% or 99% identical to any of the sequences representedby SEQ ID NOs 141, 11, 12, or(v) is selected from the group of genes having a nucleotide sequenceencoding an amino acid sequence that, when the two amino acid sequencesare optimally aligned and compared, is at least 85% preferably at least90%, 95%, 98% or 99% identical to the amino acid sequence encoded by anyof SEQ ID NOs 141, 11, 12.

In one embodiment, the present invention relates to an interferingribonucleic acid (RNA) that functions upon uptake by an insect pestspecies to down-regulate expression of a target gene in said insectpest, wherein the RNA comprises at least one silencing element whereinthe silencing element is a region of double-stranded RNA comprisingannealed complementary strands, one strand of which comprises orconsists of a sequence of nucleotides which is at least partiallycomplementary to a target nucleotide sequence within the target gene,and wherein the target gene

(i) is selected from the group of genes having a nucleotide sequencecomprising any of SEQ ID NOs 17, 18, or the complement thereof, orhaving a nucleotide sequence so that, when the two sequences areoptimally aligned and compared, is at least 75% preferably at least 80%,85%, 90%, 95%, 98% or 99% identical to any of SEQ ID NOs 17, 18, or thecomplement thereof, or(ii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 17, 18, or the complement thereof, or having a nucleotidesequence so that, when said gene comprising said fragment is optimallyaligned and compared with any of SEQ ID NOs 17, 18, said nucleotidesequence is at least 75% preferably at least 80%, 85%, 90%, 95%, 98% or99% identical to any of SEQ ID NOs 17, 18, or the complement thereof, or(iii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 17, 18, or the complement thereof, and wherein when saidfragment is optimally aligned and compared with the correspondingfragment in any of SEQ ID NOs 17, 18, said nucleotide sequence of saidfragment is at least 75% preferably at least 80%, 85%, 90%, 95%, 98% or99% identical to said corresponding fragment of any of SEQ ID NOs 17,18, or the complement thereof, or(iv) is an insect pest orthologue of a gene having a nucleotide sequencecomprising any of SEQ ID NOs 17, 18, or the complement thereof, whereinthe two orthologous genes are similar in sequence to such a degree thatwhen the two genes are optimally aligned and compared, the orthologuehas a sequence that is at least 75% preferably at least 80%, 85%, 90%,95%, 98% or 99% identical to any of the sequences represented by SEQ IDNOs 17, 18, or(v) is selected from the group of genes having a nucleotide sequenceencoding an amino acid sequence that, when the two amino acid sequencesare optimally aligned and compared, is at least 85% preferably at least90%, 95%, 98% or 99% identical to the amino acid sequence encoded by anyof SEQ ID NOs 17, 18.

In one embodiment, the present invention relates to an interferingribonucleic acid (RNA) that functions upon uptake by an insect pestspecies to down-regulate expression of a target gene in said insectpest, wherein the RNA comprises at least one silencing element whereinthe silencing element is a region of double-stranded RNA comprisingannealed complementary strands, one strand of which comprises orconsists of a sequence of nucleotides which is at least partiallycomplementary to a target nucleotide sequence within the target gene,and wherein the target gene

(i) is selected from the group of genes having a nucleotide sequencecomprising any of SEQ ID NOs 19, 20, or the complement thereof, orhaving a nucleotide sequence so that, when the two sequences areoptimally aligned and compared, is at least 75% preferably at least 80%,85%, 90%, 95%, 98% or 99% identical to any of SEQ ID NOs 19, 20, or thecomplement thereof, or(ii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 19, 20, or the complement thereof, or having a nucleotidesequence so that, when said gene comprising said fragment is optimallyaligned and compared with any of SEQ ID NOs 19, 20, said nucleotidesequence is at least 75% preferably at least 80%, 85%, 90%, 95%, 98% or99% identical to any of SEQ ID NOs 19, 20, or the complement thereof, or(iii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 19, 20, or the complement thereof, and wherein when saidfragment is optimally aligned and compared with the correspondingfragment in any of SEQ ID NOs 19, 20, said nucleotide sequence of saidfragment is at least 75% preferably at least 80%, 85%, 90%, 95%, 98% or99% identical to said corresponding fragment of any of SEQ ID NOs 19,20, or the complement thereof, or(iv) is an insect pest orthologue of a gene having a nucleotide sequencecomprising any of SEQ ID NOs 19, 20, or the complement thereof, whereinthe two orthologous genes are similar in sequence to such a degree thatwhen the two genes are optimally aligned and compared, the orthologuehas a sequence that is at least 75% preferably at least 80%, 85%, 90%,95%, 98% or 99% identical to any of the sequences represented by SEQ IDNOs 19, 20, or(v) is selected from the group of genes having a nucleotide sequenceencoding an amino acid sequence that, when the two amino acid sequencesare optimally aligned and compared, is at least 85% preferably at least90%, 95%, 98% or 99% identical to the amino acid sequence encoded by anyof SEQ ID NOs 19, 20.

In one embodiment, the present invention relates to an interferingribonucleic acid (RNA) that functions upon uptake by an insect pestspecies to down-regulate expression of a target gene in said insectpest, wherein the RNA comprises at least one silencing element whereinthe silencing element is a region of double-stranded RNA comprisingannealed complementary strands, one strand of which comprises orconsists of a sequence of nucleotides which is at least partiallycomplementary to a target nucleotide sequence within the target gene,and wherein the target gene

(i) is selected from the group of genes having a nucleotide sequencecomprising any of SEQ ID NOs 165, 166, 167, or the complement thereof,or having a nucleotide sequence so that, when the two sequences areoptimally aligned and compared, is at least 75% preferably at least 80%,85%, 90%, 95%, 98% or 99% identical to any of SEQ ID NOs 165, 166, 167,or the complement thereof, or(ii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 165, 166, 167, or the complement thereof, or having anucleotide sequence so that, when said gene comprising said fragment isoptimally aligned and compared with any of SEQ ID NOs 165, 166, 167,said nucleotide sequence is at least 75% preferably at least 80%, 85%,90%, 95%, 98% or 99% identical to any of SEQ ID NOs 165, 166, 167, orthe complement thereof, or(iii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 165, 166, 167, or the complement thereof, and wherein whensaid fragment is optimally aligned and compared with the correspondingfragment in any of SEQ ID NOs 17, 18, said nucleotide sequence of saidfragment is at least 75% preferably at least 80%, 85%, 90%, 95%, 98% or99% identical to said corresponding fragment of any of SEQ ID NOs 165,166, 167, or the complement thereof, or(iv) is an insect pest orthologue of a gene having a nucleotide sequencecomprising any of SEQ ID NOs 165, 166, 167, or the complement thereof,wherein the two orthologous genes are similar in sequence to such adegree that when the two genes are optimally aligned and compared, theorthologue has a sequence that is at least 75% preferably at least 80%,85%, 90%, 95%, 98% or 99% identical to any of the sequences representedby SEQ ID NOs 165, 166, 167, or(v) is selected from the group of genes having a nucleotide sequenceencoding an amino acid sequence that, when the two amino acid sequencesare optimally aligned and compared, is at least 85% preferably at least90%, 95%, 98% or 99% identical to the amino acid sequence encoded by anyof SEQ ID NOs 165, 166, 167.

In one embodiment, the present invention relates to an interferingribonucleic acid (RNA) that functions upon uptake by an insect pestspecies to down-regulate expression of a target gene in said insectpest, wherein the RNA comprises at least one silencing element whereinthe silencing element is a region of double-stranded RNA comprisingannealed complementary strands, one strand of which comprises orconsists of a sequence of nucleotides which is at least partiallycomplementary to a target nucleotide sequence within the target gene,and wherein the target gene

(i) is selected from the group of genes having a nucleotide sequencecomprising any of SEQ ID NOs 143, 121, 142, 176, 182, 130, 177, 183, orthe complement thereof, or having a nucleotide sequence so that, whenthe two sequences are optimally aligned and compared, is at least 75%preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to any ofSEQ ID NOs 143, 121, 142, 176, 182, 130, 177, 183, or the complementthereof, or(ii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 143, 121, 142, 176, 182, 130, 177, 183, or the complementthereof, or having a nucleotide sequence so that, when said genecomprising said fragment is optimally aligned and compared with any ofSEQ ID NOs 143, 121, 142, 176, 182, 130, 177, 183, said nucleotidesequence is at least 75% preferably at least 80%, 85%, 90%, 95%, 98% or99% identical to any of SEQ ID NOs 143, 121, 142, 176, 182, 130, 177,183, or the complement thereof, or(iii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 143, 121, 142, 176, 182, 130, 177, 183, or the complementthereof, and wherein when said fragment is optimally aligned andcompared with the corresponding fragment in any of SEQ ID NOs 143, 121,142, 176, 182, 130, 177, 183, said nucleotide sequence of said fragmentis at least 75% preferably at least 80%, 85%, 90%, 95%, 98% or 99%identical to said corresponding fragment of any of SEQ ID NOs 143, 121,142, 176, 182, 130, 177, 183, or the complement thereof, or(iv) is an insect pest orthologue of a gene having a nucleotide sequencecomprising any of SEQ ID NOs 143, 121, 142, 176, 182, 130, 177, 183, orthe complement thereof, wherein the two orthologous genes are similar insequence to such a degree that when the two genes are optimally alignedand compared, the orthologue has a sequence that is at least 75%preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to any ofthe sequences represented by SEQ ID NOs 143, 121, 142, 176, 182, 130,177, 183, or(v) is selected from the group of genes having a nucleotide sequenceencoding an amino acid sequence that, when the two amino acid sequencesare optimally aligned and compared, is at least 85% preferably at least90%, 95%, 98% or 99% identical to the amino acid sequence encoded by anyof SEQ ID NOs 143, 121, 142, 176, 182, 130, 177, 183.

In one embodiment, the present invention relates to an interferingribonucleic acid (RNA) that functions upon uptake by an insect pestspecies to down-regulate expression of a target gene in said insectpest, wherein the RNA comprises at least one silencing element whereinthe silencing element is a region of double-stranded RNA comprisingannealed complementary strands, one strand of which comprises orconsists of a sequence of nucleotides which is at least partiallycomplementary to a target nucleotide sequence within the target gene,and wherein the target gene

(i) is selected from the group of genes having a nucleotide sequencecomprising any of SEQ ID NOs 145, 122, 144, 178, 131, 179 or thecomplement thereof, or having a nucleotide sequence so that, when thetwo sequences are optimally aligned and compared, is at least 75%preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to any ofSEQ ID NOs 145, 122, 144, 178, 131, 179, or the complement thereof, or(ii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 145, 122, 144, 178, 131, 179, or the complement thereof, orhaving a nucleotide sequence so that, when said gene comprising saidfragment is optimally aligned and compared with any of SEQ ID NOs 145,122, 144, 178, 131, 179, said nucleotide sequence is at least 75%preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to any ofSEQ ID NOs 145, 122, 144, 178, 131, 179, or the complement thereof, or(iii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 145, 122, 144, 178, 131, 179, or the complement thereof, andwherein when said fragment is optimally aligned and compared with thecorresponding fragment in any of SEQ ID NOs 145, 122, 144, 178, 131,179, said nucleotide sequence of said fragment is at least 75%preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to saidcorresponding fragment of any of SEQ ID NOs 145, 122, 144, 178, 131,179, or the complement thereof, or(iv) is an insect pest orthologue of a gene having a nucleotide sequencecomprising any of SEQ ID NOs 145, 122, 144, 178, 131, 179, or thecomplement thereof, wherein the two orthologous genes are similar insequence to such a degree that when the two genes are optimally alignedand compared, the orthologue has a sequence that is at least 75%preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical to any ofthe sequences represented by SEQ ID NOs 145, 122, 144, 178, 131, 179, or(v) is selected from the group of genes having a nucleotide sequenceencoding an amino acid sequence that, when the two amino acid sequencesare optimally aligned and compared, is at least 85% preferably at least90%, 95%, 98% or 99% identical to the amino acid sequence encoded by anyof SEQ ID NOs 145, 122, 144, 178, 131, 179.

In one embodiment, the present invention relates to an interferingribonucleic acid (RNA) that functions upon uptake by an insect pestspecies to down-regulate expression of a target gene in said insectpest, wherein the RNA comprises at least one silencing element whereinthe silencing element is a region of double-stranded RNA comprisingannealed complementary strands, one strand of which comprises orconsists of a sequence of nucleotides which is at least partiallycomplementary to a target nucleotide sequence within the target gene,and wherein the target gene

(i) is selected from the group of genes having a nucleotide sequencecomprising any of SEQ ID NOs 128, 149, 184, 137, or the complementthereof, or having a nucleotide sequence so that, when the two sequencesare optimally aligned and compared, is at least 75% preferably at least80%, 85%, 90%, 95%, 98% or 99% identical to any of SEQ ID NOs 128, 149,184, 137, or the complement thereof, or(ii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 128, 149, 184, 137, or the complement thereof, or having anucleotide sequence so that, when said gene comprising said fragment isoptimally aligned and compared with any of SEQ ID NOs 128, 149, 184,137, said nucleotide sequence is at least 75% preferably at least 80%,85%, 90%, 95%, 98% or 99% identical to any of SEQ ID NOs 128, 149, 184,137, or the complement thereof, or(iii) is selected from the group of genes having a nucleotide sequencecomprising a fragment of at least 21 contiguous nucleotides of any ofSEQ ID NOs 128, 149, 184, 137, or the complement thereof, and whereinwhen said fragment is optimally aligned and compared with thecorresponding fragment in any of SEQ ID NOs 128, 149, 184, 137, saidnucleotide sequence of said fragment is at least 75% preferably at least80%, 85%, 90%, 95%, 98% or 99% identical to said corresponding fragmentof any of SEQ ID NOs 128, 149, 184, 137, or the complement thereof, or(iv) is an insect pest orthologue of a gene having a nucleotide sequencecomprising any of SEQ ID NOs 128, 149, 184, 137, or the complementthereof, wherein the two orthologous genes are similar in sequence tosuch a degree that when the two genes are optimally aligned andcompared, the orthologue has a sequence that is at least 75% preferablyat least 80%, 85%, 90%, 95%, 98% or 99% identical to any of thesequences represented by SEQ ID NOs 128, 149, 184, 137, or(v) is selected from the group of genes having a nucleotide sequenceencoding an amino acid sequence that, when the two amino acid sequencesare optimally aligned and compared, is at least 85% preferably at least90%, 95%, 98% or 99% identical to the amino acid sequence encoded by anyof SEQ ID NOs 128, 149, 184, 137.

In yet other embodiments, the present invention relates to aninterfering ribonucleic acid (RNA or double stranded RNA) that inhibitsor downregulates the expression of a target gene that encodes amitochondrial cytochrome c oxidase subunit II protein (e.g. an insectorthologue of the CG34069 Dm protein).

Thus, in one aspect, the invention provides an interfering ribonucleicacid (RNA) that functions upon uptake by an insect pest species todown-regulate expression of a target gene in said insect pest.

As used herein, a “target gene” comprises any gene in the insect pestwhich one intends to down-regulate. In a preferred embodiment, thetarget gene is down-regulated so as to control pest infestation, forexample by disrupting an essential biological process occurring in thepest, or by decreasing the pathogenicity of the pest. Preferred targetgenes therefore include but are not limited to those that play key rolesin regulating feeding, survival, growth, development, reproduction,infestation and infectivity. According to one embodiment, the targetgene is such that when its expression is down-regulated or inhibited,the insect pest is killed. According to another embodiment, the targetgene is such that when its expression is down-regulated or inhibited,growth of the pest is prevented or retarded or stunted or delayed orimpeded, pest reproduction is prevented, or transition through the lifecycles of the pest is prevented. According to yet another embodiment ofthe invention, the target gene is such that when its expression isdown-regulated or inhibited, the damage caused by the pest and/or theability of the pest to infect or infest environments, surfaces and/orplant or crop species is reduced; or the pest stops feeding from itsnatural food resources such as plants and plant products. The terms“infest” and “infect” or “infestation” and “infection” are generallyused interchangeably throughout.

The target genes may be expressed in all or some of the cells of theinsect pest. Furthermore, the target genes may only be expressed by theinsect pest at a particular stage of its life-cycle, for example, themature adult phase, immature nymph or larval phase or egg phase.

As used herein “pest” species are preferably insect species that causeinfection or infestation, preferably of plants. The insect species maycomprise and species belonging to the Orders Coleoptera, Lepidoptera,Diptera, Dichyoptera, Orthoptera, Hemiptera, or Siphonaptera.

Preferred plant pathogenic insects according to the invention are plantpest are selected from the group consisting of Leptinotarsa spp. (e.g.L. decemlineata (Colorado potato beetle), L. juncta (false potatobeetle), or L. texana (Texan false potato beetle)); Nilaparvata spp.(e.g. N. lugens (brown planthopper)); Laodelphax spp. (e.g. L.striatellus (small brown planthopper)); Nephotettix spp. (e.g. N.virescens or N. cincticeps (green leafhopper), or N. nigropictus (riceleafhopper)); Sogatella spp. (e.g. S. furcifera (white-backedplanthopper)); Chilo spp. (e.g. C. suppressalis (rice striped stemborer), C. auricilius (gold-fringed stem borer), or C. polychrysus(dark-headed stem borer)); Sesamia spp. (e.g. S. inferens (pink riceborer)); Tryporyza spp. (e.g. T. innotata (white rice borer), or T.incertulas (yellow rice borer)); Anthonomus spp. (e.g. A. grandis (bollweevil)); Phaedon spp. (e.g. P. cochleariae (mustard leaf beetle));Epilachna spp. (e.g. E. varivetis (mexican bean beetle)); Tribolium spp.(e.g. T. castaneum (red floor beetle)); Diabrotica spp. (e.g. D.virgifera virgifera (western corn rootworm), D. barberi (northern cornrootworm), D. undecimpunctata howardi (southern corn rootworm), D.virgifera zeae (Mexican corn rootworm); Ostrinia spp. (e.g. O. nubilalis(European corn borer)); Anaphothrips spp. (e.g. A. obscrurus (grassthrips)); Pectinophora spp. (e.g. P. gossypiella (pink bollworm));Heliothis spp. (e.g. H. virescens (tobacco budworm)); Trialeurodes spp.(e.g. T. abutiloneus (banded-winged whitefly) T. vaporariorum(greenhouse whitefly)); Bemisia spp. (e.g. B. argentifolii (silverleafwhitefly)); Aphis spp. (e.g. A. gossypii (cotton aphid)); Lygus spp.(e.g. L. lineolaris (tarnished plant bug) or L. hesperus (westerntarnished plant bug)); Euschistus spp. (e.g. E. conspersus (conspersestink bug)); Chlorochroa spp. (e.g. C. sayi (Say stinkbug)); Nezara spp.(e.g. N. viridula (southern green stinkbug)); Thrips spp. (e.g. T.tabaci (onion thrips)); Frankliniella spp. (e.g. F. fusca (tobaccothrips), or F. occidentalis (western flower thrips)); Acheta spp. (e.g.A. domesticus (house cricket)); Myzus spp. (e.g. M. persicae (greenpeach aphid)); Macrosiphum spp. (e.g. M. euphorbiae (potato aphid));Blissus spp. (e.g. B. leucopterus leucopterus (chinch bug)); Acrosternumspp. (e.g. A. hilare (green stink bug)); Chilotraea spp. (e.g. C.polychrysa (rice stalk borer)); Lissorhoptrus spp. (e.g. L. oryzophilus(rice water weevil)); Rhopalosiphum spp. (e.g. R. maidis (corn leafaphid)); and Anuraphis spp. (e.g. A. maidiradicis (corn root aphid)).

According to a more specific embodiment, the invention is applicable forspecies belonging to the family of Chrysomelidae or leaf beetles.Chrysomelid beetles such Colorado potato Beetles, Flea Beetles, CornRootworms and Curculionids such as Alfalfa Weevils are particularlyimportant pests. Specific Leptinotarsa species to control according tothe invention include Colorado Potato Beetle (Leptinotarsa decemlineata(Say) and False Potato Beetle (Leptinotarsa juncta (Say). CPB is a(serious) pest on our domestic potato, other cultivated and wild tuberbearing and non-tuber bearing potato species and other Solanaceous(nightshades) plant species including the crop species tomato, eggplant,peppers, tobacco (Nicotiana species including ornamentals), groundcherry, rice, corn or cotton; and the weed/herb species, horse nettle,common nightshade, thorn apple, henbane and buffalo burr. Corn rootwormsinclude species found in the genus Diabrotica (e.g., D. undecimpunctataundecimpunctata, D. undecimpunctata howardii, D. longicomis, D.virgifera and D. balteata). Corn rootworms cause extensive damage tocorn and curcubits.

According to a more specific embodiment, the invention is applicable forspecies belonging to the order of Hemipterans (family of Aphidoidea),such as Myzus persicae (green peach aphid, Aphis fabae (Black BeanAphid), Acyrthosiphum pisum (Pea Aphid), Brevicoryne brassicae (CabbageAphid), Sitobion avenae (Grain Aphid), Cavariella aegopodii (CarrotAphid), Aphis craccivora (Groundnut Aphid), Aphis gossypii (CottonAphid), Toxoptera aurantii (Black Citrus Aphid), Cavariella spp (WillowAphid), Chaitophorus spp (Willow Leaf Aphids), Cinara spp. (Black PineAphids), Drepanosiphum platanoides (Sycamore Aphid) Elatobium spp(Spruce Aphids) which cause damage to plants such as Prunus trees,particularly peach, apricot and plum; trees that are mainly cultured forwood production such as willows and poplars, to row crops such as corn,cotton, soy, wheat and rice, to vegetable crops of the familiesSolanaceae, Chenopodiaceae, Compositae, Cruciferae, and Cucurbitaceae,including but not limited to, artichoke, asparagus, bean, beets,broccoli, Brussels sprouts, cabbage, carrot, cauliflower, cantaloupe,celery, corn, cucumber, fennel, kale, kohlrabi, turnip, eggplant,lettuce, mustard, okra, parsley, parsnip, pea, pepper, potato, radish,spinach, squash, tomato, turnip, watercress, and watermelon; or fieldcrops such as, but not limited to, tobacco, sugar beet, and sunflower; aflower crop or other ornamental plant such as pine trees and conifers.Other Hemipterans belong to Nilaparvata ssp (eg. N. lugens, Sogatellafurcifera) and cause damage to rice plants. Other Hemipterans belong toLygus ssp (eg. Lygus hesperus, Lygus rugulipennis, Lygus lineolaris,Lygus sully) and other species of plant-feeding insects in the family ofthe Miridae, and cause damage to cotton, potato plants, strawberries,cotton, alfalfa, canola, peach, plums, grape, lettuce, eggplant, onion,green beans. As well as several Mediterranean trees and severalornamental trees such as elm tree (Ulmus spp.) pine nut (Pinus Pinea)London plane tree (Platanus Acerifolia), white redbud (Malus alba).Other Hemipterans belong to the family of the Pentatomoidea, they arecommonly referred to as shield bugs, chust bugs, and stink bugs (eg; thebrown marmorated stink bug (Halyomorpha halys), the Consperse stink bug(Euschistus conspersus), southern green stink bug (Nezara viridula),forest bug (Pentatoma rufipes), harlequin bug (Murgantia histrionica),rice stink bug (Oebalus pugnax)) and cause damage to fruits includingapples, peaches, figs, mulberries, citrus fruits and persimmons,blackberry, and vegetables including sweetcorn, tomatoes, soy beans,lima beans and green peppers, cabbage, cauliflower, turnips,horseradish, collards, mustard, Brussels sprouts, potato, egg plant,okra, beans, asparagus, beets, weeds, fruit trees and field crops suchas field corn and soy bean. Stink bugs are also a pest of grasses,sorghum and rice.

A plant to be used in the methods of the invention, or a transgenicplant according to the invention encompasses any plant, but ispreferably a plant that is susceptible to infestation by a plantpathogenic insect.

Accordingly, the present invention extends to plants and to methods asdescribed herein wherein the plant is chosen from the following group ofplants (or crops): alfalfa, apple, apricot, artichoke, asparagus,avocado, banana, barley, beans, beet, blackberry, blueberry, broccoli,Brussels sprouts, cabbage, canola, carrot, cassava, cauliflower, acereal, celery, cherry, citrus, clementine, coffee, corn, cotton,cucumber, eggplant, endive, eucalyptus, figs, grape, grapefruit,groundnuts, ground cherry, kiwifruit, lettuce, leek, lemon, lime, pine,maize, mango, melon, millet, mushroom, nut oat, okra, onion, orange, anornamental plant or flower or tree, papaya, parsley, pea, peach, peanut,peat, pepper, persimmon, pineapple, plantain, plum, pomegranate, potato,pumpkin, radicchio, radish, rapeseed, raspberry, rice, rye, sorghum,soy, soybean, spinach, strawberry, sugar beet, sugarcane, sunflower,sweet potato, tangerine, tea, tobacco, tomato, a vine, watermelon,wheat, yams and zucchini.

In specific embodiments, the present invention provides target geneswhich encode proteins involved in the function of a wings up A (troponinI), a mitochondrial cytochrome c oxidase subunit II protein, or one ofthe ribosomal proteins as specified in Table 1.

In preferred embodiments, the present invention provides target genesselected from the group of genes (i) having a nucleotide sequencecomprising any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13,14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62,19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78,143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135,226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153,246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389,or the complement thereof, or having a nucleotide sequence so that, whenthe two sequences are optimally aligned and compared, is at least 75%,preferably at least 80%, 85%, 90%, 95%, 98% or 99% identical any of SEQID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285,294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8,39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15,204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66,21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142,176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122,144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124,133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229,127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302 to305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to 249,154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160,161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273,168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to 317,186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, or thecomplement thereof, or (ii) having a nucleotide sequence consisting ofany of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38,140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20,63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143,121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301,145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237,302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, or (iii) having a nucleotide sequence comprisinga fragment of at least 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35,40, 45, 50, 55, 60, 70, 80, 90, 100, 110, 125, 150, 175, 200, 225, 250,300, 350, 400, 450, 500, 550, 600, 700, 800, 900 1000, 1100, 1200, 1300,1400, 1500, 2000, or 3000 contiguous nucleotides of any of SEQ ID NOs 1,174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297,310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9,10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205,55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142, 176, 182, 130,177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131,179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221,146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230to 233, 128, 149, 184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238to 241, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265,163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273, 168, 170, 169, 274to 277, 172, 173, 278 to 281, 200, 201, 314 to 317, 186, 202, 187, 203,306 to 309, 318 to 321, 386, 387, 388, 389, or the complement thereof,or having a nucleotide sequence so that, when said gene comprising saidfragment is optimally aligned and compared with any of SEQ ID NOs 1,174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297,310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9,10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205,55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142, 176, 182, 130,177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131,179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221,146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230to 233, 128, 149, 184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238to 241, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265,163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273, 168, 170, 169, 274to 277, 172, 173, 278 to 281, 200, 201, 314 to 317, 186, 202, 187, 203,306 to 309, 318 to 321, 386, 387, 388, 389, said nucleotide sequence isat least 75% preferably at least 80%, 85%, 90%, 95%, 98% or 99%identical to any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189,27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6,35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50,13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132,214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126,135, 226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234to 237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152,153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159,258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167,166, 270 to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200,201, 314 to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387,388, 389, or the complement thereof, or (iv) having a nucleotidesequence comprising a fragment of at least 20, 21, 22, 23, 24, 25, 26,27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 110, 125, 150,175, 200, 225, 250, 300, 350, 400, 450, 500, 550, 600, 700, 800, 9001000, 1100, 1200, 1300, 1400, 1500, 2000, or 3000 contiguous nucleotidesof any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30,282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38,140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20,63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143,121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301,145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237,302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, and wherein when said fragment is optimallyaligned and compared with the corresponding fragment in any of SEQ IDNOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15,204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66,21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142,176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122,144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124,133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229,127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302 to305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to 249,154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160,161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273,168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to 317,186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, saidnucleotide sequence of said fragment is at least 75% preferably at least80%, 85%, 90%, 95%, 98% or 99% identical to said corresponding fragmentof any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30,282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38,140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20,63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143,121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301,145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237,302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, or (v) having a nucleotide sequence encoding anamino acid sequence that, when the two amino acid sequences areoptimally aligned and compared, is at least 70% preferably at least 75%,80%, 85%, 90%, 95%, 98% or 99% identical to the amino acid sequenceencoded by any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13,14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62,19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78,143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135,226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153,246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389or (vi) which gene is an insect pest orthologue of a gene having anucleotide sequence comprising any of SEQ ID NOs 1, 174, 404, 180, 188,2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141,11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to74, 25, 26, 75 to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128,149, 184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150,151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157,254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164,266 to 269, 165, 167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172,173, 278 to 281, 200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309,318 to 321, 386, 387, 388, 389, or the complement thereof, wherein thetwo orthologous genes are similar in sequence to such a degree that whenthe two genes are optimally aligned and compared, the orthologue has asequence that is at least 75% preferably at least 80%, 85%, 90%, 95%,98% or 99% identical to any of the sequences represented by SEQ ID NOs1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204,16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21,22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142, 176,182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144,178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133,218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127,148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302 to 305,129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to 249, 154,155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161,262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273, 168,170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to 317, 186,202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389;

and wherein the nucleotide sequence of said gene is no longer than10000, 9000, 8000, 7000, 6000, 5000, 4000, 3000, 2000 or 1500nucleotides.

The amino acid sequences encoded by the target genes of the presentinvention are represented by SEQ ID NOs. SEQ ID NOs 79, 349, 405, 352,356, 80, 326, 81, 327, 82, 83, 328, 84, 329, 85, 86, 359, 87 to 91, 330,350, 353, 331, 351, 332 to 336, 337, 354, 338 to 344, 346, 345, 347,348, 357, 355, 358, 390 to 393.

As used herein, the term “having” has the same meaning as “comprising”.

As used herein, the term “sequence identity” is used to describe thesequence relationship between two or more nucleotide or amino acidsequences. The percentage of “sequence identity” between two sequencesis determined by comparing two optimally aligned sequences over acomparison window (a defined number of positions), wherein the portionof the sequence in the comparison window may comprise additions ordeletions (i.e. gaps) as compared to the reference sequence in order toachieve optimal alignment. The percentage sequence identity iscalculated by determining the number of positions at which the identicalnucleotide base or amino acid residue occurs in both sequences to yieldthe number of ‘matched’ positions, dividing the number of matchedpositions by the total number of positions in the comparison window andmultiplying the result by 100. Methods and software for determiningsequence identity are available in the art and include the Blastsoftware and GAP analysis. For nucleic acids, the percent identity iscalculated preferably by the BlastN alignment tool whereby the percentidentity is calculated over the entire length of the query nucleotidesequence. A person skilled in the art will recognise that homologues ororthologues (homologues existing in different species) of the targetgenes represented by any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175,181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34,139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12,47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17,18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25,26, 75 to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293,123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225,147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184,137, 185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257,158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269,165, 167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to281, 200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309, 318 to 321,386, 387, 388, 389 can be identified. These pest homologues and/ororthologues are also within the scope of the current invention.Preferred homologues and/or orthologues are genes similar in nucleotidesequence to such a degree that when the two genes are optimally alignedand compared, the homologue and/or orthologue has a sequence that is atleast 75%, preferably at least 80% or 85%, more preferably at least 90%or 95%, and most preferably at least about 99% identical to any of SEQID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285,294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8,39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15,204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66,21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142,176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122,144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124,133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229,127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302 to305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to 249,154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160,161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273,168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to 317,186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389 or thecomplement thereof. Similarly, also preferred homologues and/ororthologues are proteins that are similar in amino acid sequence to sucha degree that when the two amino acid sequences are optimally alignedand compared, the homologue and/or orthologue has a sequence that is atleast 75%, preferably at least 80% or 85%, more preferably at least 90%or 95%, and most preferably at least about 99% identical to any of SEQID NOs 79, 349, 405, 352, 356, 80, 326, 81, 327, 82, 83, 328, 84, 329,85, 86, 359, 87 to 91, 330, 350, 353, 331, 351, 332 to 336, 337, 354,338 to 344, 346, 345, 347, 348, 357, 355, 358, 390 to 393.

Other homologues are genes which are alleles of a gene comprising asequence as represented by any of SEQ ID NOs 1, 174, 404, 180, 188, 2,175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11,12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325,17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74,25, 26, 75 to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to 209,286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128, 149,184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150, 151,242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to269, 165, 167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172, 173, 278to 281, 200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309, 318 to321, 386, 387, 388, 389. Further preferred homologues are genescomprising at least one single nucleotide polymorphism (SNP) compared toa gene comprising a sequence as represented by any of SEQ ID NOs 1, 174,404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10,43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70,23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142, 176, 182, 130, 177,183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179,210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146,125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to233, 128, 149, 184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238 to241, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253,156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163,162, 164, 266 to 269, 165, 167, 166, 270 to 273, 168, 170, 169, 274 to277, 172, 173, 278 to 281, 200, 201, 314 to 317, 186, 202, 187, 203, 306to 309, 318 to 321, 386, 387, 388, 389.

The ‘interfering ribonucleic acid (RNA)’ of the current inventionencompasses any type of RNA molecule capable of down-regulating or‘silencing’ expression of a target gene, including but not limited tosense RNA, antisense RNA, short interfering RNA (sRNA), microRNA(miRNA), double-stranded RNA (dsRNA), hairpin RNA (RNA) and the like.Methods to assay for functional interfering RNA molecules are well knownin the art and are disclosed elsewhere herein.

The interfering RNA molecules of the current invention effectsequence-specific down-regulation of expression of a target gene bybinding to a target nucleotide sequence within the target gene. Bindingoccurs as a result of base pairing between complementary regions of theinterfering RNA and the target nucleotide sequence. As used herein, theterm ‘silencing element’ refers to the portion or region of theinterfering RNA comprising or consisting of a sequence of nucleotideswhich is complementary, or at least partially complementary, to a targetnucleotide sequence within the target gene, and which functions as theactive portion of the interfering RNA to direct down-regulation ofexpression of said target gene. In one embodiment of the invention, thesilencing element comprises or consists of a sequence of at least 17contiguous nucleotides, preferably at least 18 or 19 contiguousnucleotides, more preferably at least 21 contiguous nucleotides, evenmore preferably at least 22, 23, 24 or 25 contiguous nucleotidescomplementary to a target nucleotide sequence within the target gene.

As used herein, “expression of a target gene” refers to thetranscription and accumulation of the RNA transcript encoded by a targetgene and/or translation of the mRNA into protein. The term‘down-regulate’ is intended to refer to any of the methods known in theart by which interfering RNA molecules reduce the level of primary RNAtranscripts, mRNA or protein produced from a target gene. In certainembodiments, down-regulation refers to a situation whereby the level ofRNA or protein produced from a gene is reduced by at least 10%,preferably by at least 33%, more preferably by at least 50%, yet morepreferably by at least 80%. In particularly preferred embodiments,down-regulation refers to a reduction in the level of RNA or proteinproduced from a gene by at least 80%, preferably by at least 90%, morepreferably by at least 95%, and most preferably by at least 99% withincells of the insect pest as compared with an appropriate control insectpest which has for example, not been exposed to an interfering RNA orhas been exposed to a control interfering RNA molecule. Methods fordetecting reductions in RNA or protein levels are well known in the artand include RNA solution hybridization, Northern hybridization, reversetranscription (e.g. quantitative RT-PCR analysis), microarray analysis,antibody binding, enzyme-linked immunosorbent assay (ELISA) and Westernblotting. In another embodiment of the invention, down-regulation refersto a reduction in RNA or protein levels sufficient to result in adetectable change in a phenotype of the pest as compared with anappropriate pest control, for example, cell death, cessation of growth,or the like. Down-regulation can thus be measured by phenotypic analysisof the insect pest using techniques routine in the art.

In a preferred embodiment of the invention, the interfering RNAdown-regulates gene expression by RNA interference or RNAi. RNAi is aprocess of sequence-specific gene regulation typically mediated bydouble-stranded RNA molecules such as short interfering RNAs (siRNAs).siRNAs comprise a sense RNA strand annealed by complementary basepairingto an antisense RNA strand. The sense strand or ‘guide strand’ of thesiRNA molecule comprises a sequence of nucleotides complementary to asequence of nucleotides located within the RNA transcript of the targetgene. The sense strand of the siRNA is therefore able to anneal to theRNA transcript via Watson-Crick-type basepairing and target the RNA fordegradation within a cellular complex known as the RNAi-inducedsilencing complex or RISC. Thus, in the context of preferred interferingRNA molecules of the current invention, the silencing element asreferred to herein may be a double-stranded region comprising annealedcomplementary strands, at least one strand of which comprises orconsists of a sequence of nucleotides which is complementary or at leastpartially complementary to a target nucleotide sequence within a targetgene. In one embodiment the double-stranded region has a length of atleast 21, 22, 23, 24, 25, 30, 35, 40, 50, 55, 60, 70, 80, 90, 100, 125,150, 175, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200,1300, 1400, 1500, 2000 or 3000 base pairs.

Longer double-stranded RNA (dsRNA) molecules comprising one or morefunctional double-stranded silencing elements as described elsewhereherein, and capable of RNAi-mediated gene silencing are alsocontemplated within the scope of the current invention. Such longerdsRNA molecules comprise at least 80, 200, 300, 350, 400, 450, 500, 550,600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 2000 or 3000base pairs. These dsRNA molecules may serve as precursors for the activesiRNA molecules that direct the RNA transcript to the RISC complex forsubsequent degradation. dsRNA molecules present in the environmentsurrounding an organism or the cells thereof may be taken up by theorganism and processed by an enzyme called Dicer to yield siRNAmolecules. Alternatively, the dsRNA may be produced in vivo i.e.transcribed from a polynucleotide or polynucleotides encoding the samepresent within a cell, for instance a bacterial cell or a plant cell,and subsequently processed by Dicer either within the host cell orpreferably within the insect pest cells following uptake of the longerprecursor dsRNA. The dsRNA may be formed from two separate (sense andantisense) RNA strands that anneal by virtue of complementary basepairing. Alternatively, the dsRNA may be a single strand that is capableof folding back on itself to form a hairpin RNA (RNA) or stem-loopstructure. In the case of a RNA, the double-stranded region or ‘stem’ isformed from two regions or segments of the RNA that are essentiallyinverted repeats of one another and possess sufficient complementarityto allow the formation of a double-stranded region. One or morefunctional double-stranded silencing elements may be present in this‘stem region’ of the molecule. The inverted repeat regions are typicallyseparated by a region or segment of the RNA known as the ‘loop’ region.This region can comprise any nucleotide sequence conferring enoughflexibility to allow self-pairing to occur between the flankingcomplementary regions of the RNA. In general, the loop region issubstantially single-stranded and acts as a spacer element between theinverted repeats.

All the interfering RNA molecules of the invention effectsequence-specific down-regulation of expression of a target gene bybinding to a target nucleotide sequence within the target gene. Bindingoccurs as a result of complementary base pairing between the silencingelement of the interfering RNA and the target nucleotide sequence. Theinterfering RNA molecules of the invention comprise at least one or atleast two silencing elements. In one embodiment of the currentinvention, the target nucleotide sequence comprises a sequence ofnucleotides as represented by the RNA transcript of the target gene, ora fragment thereof wherein the fragment is preferably at least 17nucleotides, more preferably at least 18, 19 or 20 nucleotides, or mostpreferably at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45,50, 55, 60, 70, 80, 90, 100, 110, 125, 150, 175, 200, 225, 250, 300,350, 400, 450, 500, 550, 600, 700, 800, 900, 1000, 1100, 1200, 1300,1400, 1500, 2000 or 3000 nucleotides. In a preferred embodiment of thecurrent invention, the target nucleotide sequence comprises a sequenceof nucleotides equivalent to the RNA transcript encoded by any of thepolynucleotides selected from the group consisting of (i) apolynucleotide which comprises at least 21, preferably at least 22, 23,24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100,110, 125, 150, 175, 200, 225, 250, 300, 350, 400, 450, 500, 550, 600,700, 800, 900, 1000, 1100 or 1115 contiguous nucleotides of a nucleotidesequence as represented by any of SEQ ID NOs 1, 174, 404, 180, 188, 2,175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11,12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325,17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74,25, 26, 75 to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to 209,286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128, 149,184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150, 151,242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to269, 165, 167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172, 173, 278to 281, 200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309, 318 to321, 386, 387, 388, 389, or the complement thereof, or (ii) apolynucleotide which consists of at least 21, preferably at least 22,23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100,110, 125, 150, 175, 200, 225, 250, 300, 350, 400, 450, 500, 550, 600,700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 2000 or 3000contiguous nucleotides of a nucleotide sequence as represented by any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof,

or (iii) a polynucleotide which comprises at least 21, preferably atleast 22, 23 or 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 70,80, 90, 100, 110, 125, 150, 175, 200, 225, 250, 300, 350, 400, 450, 500,550, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 2000 or3000 contiguous nucleotides of a nucleotide sequence as represented inany of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38,140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20,63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143,121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301,145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237,302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, so that, when the two sequences are optimallyaligned and compared, said polynucleotide is at least 75% preferably atleast 80%, 85%, 90%, 95%, 98% or 99% identical to any of SEQ ID NOs 1,174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297,310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9,10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205,55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142, 176, 182, 130,177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131,179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221,146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230to 233, 128, 149, 184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238to 241, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265,163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273, 168, 170, 169, 274to 277, 172, 173, 278 to 281, 200, 201, 314 to 317, 186, 202, 187, 203,306 to 309, 318 to 321, 386, 387, 388, 389, or the complement thereof,or (iv) a polynucleotide which comprises a fragment of at least 21,preferably at least 22, 23 or 24, 25, 26, 27, 28, 29, 30, 35, 40, 45,50, 55, 60, 70, 80, 90, 100, 110, 125, 150, 175, 200, 225, 250, 300,350, 400, 450, 500, 550, 600, 700, 800, 900, 1000, 1100, 1200, 1300,1400, 1500, 2000 or 3000 contiguous nucleotides of a nucleotide asrepresented in any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181,189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139,5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289,298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123,132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147,126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137,185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245,152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158,159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165,167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281,200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386,387, 388, 389, or the complement thereof, and wherein said fragment orsaid complement has a nucleotide sequence so that, when said fragment isoptimally aligned and compared with the corresponding fragment in any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389,said nucleotide sequence is at least 75% preferably at least 80%, 85%,90%, 95%, 98% or 99% identical to said corresponding fragment of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389 orthe complement thereof,or (v) a polynucleotide which consists of a fragment of at least 21,preferably at least 22, 23 or 24, 25, 26, 27, 28, 29, 30, 35, 40, 45,50, 55, 60, 70, 80, 90, 100, 110, 125, 150, 175, 200, 225, 250, 300,350, 400, 450, 500, 550, 600, 700, 800, 900, 1000, 1100, 1200, 1300,1400, 1500, 2000 or 3000 contiguous nucleotides of a nucleotide asrepresented in any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181,189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139,5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289,298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123,132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147,126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137,185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245,152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158,159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165,167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281,200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386,387, 388, 389, or the complement thereof, and wherein said fragment orsaid complement has a nucleotide sequence that, when said fragment isoptimally aligned and compared with the corresponding fragment in any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389,said nucleotide sequence is at least 75% preferably at least 80%, 85%,90%, 95%, 98% or 99% identical to said corresponding fragment of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389 orthe complement thereof, or (vi) a polynucleotide encoding an amino acidsequence that, when the two amino acid sequences are optimally alignedand compared, is at least 70% preferably at least 75%, 80%, 85%, 90%,95%, 98% or 99% identical to the amino acid sequence encoded by any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389. Ina more preferred embodiment of the above, said polynucleotide is nolonger than 10000, 9000, 8000, 7000, 6000, 5000, 4000, 3000, 2000 or1500 nucleotides.

Preferably, the interfering RNA molecules of the current inventioncomprise at least one double-stranded region, typically the silencingelement of the interfering RNA, comprising a sense RNA strand annealedby complementary basepairing to an antisense RNA strand wherein thesense strand of the dsRNA molecule comprises a sequence of nucleotidescomplementary to a sequence of nucleotides located within the RNAtranscript of the target gene.

The silencing element, or at least one strand thereof wherein thesilencing element is double-stranded, may be fully complementary orpartially complementary to the target nucleotide sequence of the targetgene. As used herein, the term “fully complementary” means that all thebases of the nucleotide sequence of the silencing element arecomplementary to or ‘match’ the bases of the target nucleotide sequence.The term “at least partially complementary” means that there is lessthan a 100% match between the bases of the silencing element and thebases of the target nucleotide sequence. The skilled person willunderstand that the silencing element need only be at least partiallycomplementary to the target nucleotide sequence in order to mediatedown-regulation of expression of the target gene. It is known in the artthat RNA sequences with insertions, deletions and mismatches relative tothe target sequence can still be effective at RNAi. According to thecurrent invention, it is preferred that the silencing element and thetarget nucleotide sequence of the target gene share at least 80% or 85%sequence identity, preferably at least 90% or 95% sequence identity, ormore preferably at least 97% or 98% sequence identity and still morepreferably at least 99% sequence identity. Alternatively, the silencingelement may comprise 1, 2 or 3 mismatches as compared with the targetnucleotide sequence over every length of 24 partially complementarynucleotides.

It will be appreciated by the person skilled in the art that the degreeof complementarity shared between the silencing element and the targetnucleotide sequence may vary depending on the target gene to bedown-regulated or depending on the insect pest species in which geneexpression is to be controlled.

In another embodiment of the current invention, the silencing elementcomprises a sequence of nucleotides that is the RNA equivalent of any ofthe polynucleotides selected from the group consisting of apolynucleotide which comprises at least 21, preferably at least 22, 23,24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100,110, 125, 150, 175, 200, 225, 250, 300, 350, 400, 450, 500, 550, 600,700, 800, 900, 1000, 1100 or 1115 contiguous nucleotides of a nucleotidesequence as represented by any of SEQ ID NOs 1, 174, 404, 180, 188, 2,175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11,12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325,17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74,25, 26, 75 to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to 209,286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128, 149,184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150, 151,242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to269, 165, 167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172, 173, 278to 281, 200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309, 318 to321, 386, 387, 388, 389, or the complement thereof, or (ii) apolynucleotide which comprises at least 21, preferably at least 22, 23or 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100,110, 125, 150, 175, 200, 225, 250, 300, 350, 400, 450, 500, 550, 600,700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 2000 or 3000contiguous nucleotides of a nucleotide sequence as represented in any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, so that, when the two sequences are optimallyaligned and compared, said polynucleotide is at least 75% preferably atleast 80%, 85%, 90%, 95%, 98% or 99% identical to any of SEQ ID NOs 1,174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297,310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9,10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205,55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142, 176, 182, 130,177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131,179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221,146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230to 233, 128, 149, 184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238to 241, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265,163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273, 168, 170, 169, 274to 277, 172, 173, 278 to 281, 200, 201, 314 to 317, 186, 202, 187, 203,306 to 309, 318 to 321, 386, 387, 388, 389, or the complement thereof,or (iii) a polynucleotide which comprises a fragment of at least 21,preferably at least 22, 23 or 24, 25, 26, 27, 28, 29, 30, 35, 40, 45,50, 55, 60, 70, 80, 90, 100, 110, 125, 150, 175, 200, 225, 250, 300,350, 400, 450, 500, 550, 600, 700, 800, 900, 1000, 1100, 1200, 1300,1400, 1500, 2000 or 3000 contiguous nucleotides of a nucleotide asrepresented in any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181,189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139,5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289,298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123,132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147,126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137,185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245,152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158,159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165,167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281,200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386,387, 388, 389, or the complement thereof, and wherein said fragment orsaid complement has a nucleotide sequence so that, when said fragment isoptimally aligned and compared with the corresponding fragment in any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389,said nucleotide sequence is at least 75% preferably at least 80%, 85%,90%, 95%, 98% or 99% identical to said corresponding fragment of any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, wherein said polynucleotide is no longer than10000, 9000, 8000, 7000, 6000, 5000, 4000, 3000, 2000 or 1500nucleotides. It will be appreciated that in such embodiments thesilencing element may comprise or consist of a region of double-strandedRNA comprising annealed complementary strands, one strand of which, thesense strand, comprises a sequence of nucleotides at least partiallycomplementary to a target nucleotide sequence within a target gene.

The target nucleotide sequence may be selected from any suitable regionor nucleotide sequence of the target gene or RNA transcript thereof. Forexample, the target nucleotide sequence may be located within the 5′UTRor 3′UTR of the target gene or RNA transcript or within exonic orintronic regions of the gene.

The skilled person will be aware of methods of identifying the mostsuitable target nucleotide sequences within the context of thefull-length target gene. For example, multiple silencing elementstargeting different regions of the target gene can be synthesised andtested. Alternatively, digestion of the RNA transcript with enzymes suchas RNAse H can be used to determine sites on the RNA that are in aconformation susceptible to gene silencing. Target sites may also beidentified using in silico approaches, for example, the use of computeralgorithms designed to predict the efficacy of gene silencing based ontargeting different sites within the full-length gene.

The interfering RNAs of the current invention may comprise one silencingelement or multiple silencing elements, wherein each silencing elementcomprises or consists of a sequence of nucleotides which is at leastpartially complementary to a target nucleotide sequence within a targetgene and that functions upon uptake by an insect pest species todown-regulate expression of said target gene. Concatemeric RNAconstructs of this type are described in WO2006/046148 as incorporatedherein by reference. In the context of the present invention, the term‘multiple’ means at least two, at least three, at least four, etc and upto at least 10, 15, 20 or at least 30. In one embodiment, theinterfering RNA comprises multiple copies of a single silencing elementi.e. repeats of a silencing element that binds to a particular targetnucleotide sequence within a specific target gene. In anotherembodiment, the silencing elements within the interfering RNA compriseor consist of different sequences of nucleotides complementary todifferent target nucleotide sequences. It should be clear thatcombinations of multiple copies of the same silencing element combinedwith silencing elements binding to different target nucleotide sequencesare within the scope of the current invention.

The different target nucleotide sequences may originate from a singletarget gene in an insect pest species in order to achieve improveddown-regulation of a specific target gene in an insect pest species. Inthis case, the silencing elements may be combined in the interfering RNAin the original order in which the target nucleotide sequences occur inthe target gene, or the silencing elements may be scrambled and combinedrandomly in any rank order in the context of the interfering RNA ascompared with the order of the target nucleotide sequences in the targetgene.

Alternatively, the different target nucleotide sequences arerepresenting a single target gene but originating from different insectpest species.

Alternatively, the different target nucleotide sequences may originatefrom different target genes. If the interfering RNA is for use inpreventing and/or controlling pest infestation, it is preferred that thedifferent target genes are chosen from the group of genes regulatingessential biological functions of insect pest species, including but notlimited to survival, growth, development, reproduction andpathogenicity. The target genes may regulate the same or differentbiological pathways or processes. In one embodiment, at least one of thesilencing elements comprises or consists of a sequence of nucleotideswhich is at least partially complementary to a target nucleotidesequence within a target gene wherein the target gene is selected fromthe group of genes as described earlier.

In a further embodiment of the invention, the different genes targetedby the different silencing elements originate from the same insect pestspecies. This approach is designed to achieve enhanced attack against asingle insect pest species. In particular, the different target genesmay be expressed differentially in the different stages of the insect'slife cycle, for example, the mature adult, immature larval and eggstages. The interfering RNA of the invention may thus be used to preventand/or control insect pest infestation at more than one stage of theinsect's life cycle.

In an alternative embodiment of the invention, the different genestargeted by the different silencing elements originate from differentinsect pest species. The interfering RNA of the invention can thus beused to prevent and/or control infestation by more than one insect pestspecies simultaneously. The silencing elements may be arranged as onecontiguous region of the interfering RNA or may be separated by thepresence of linker sequences. The linker sequence may comprise a shortrandom nucleotide sequence that is not complementary to any targetnucleotide sequences or target genes. In one embodiment, the linker is aconditionally self-cleaving RNA sequence, preferably a pH-sensitivelinker or a hydrophobic-sensitive linker. In one embodiment, the linkercomprises a sequence of nucleotides equivalent to an intronic sequence.Linker sequences of the current invention may range in length from about1 base pair to about 10000 base pairs, provided that the linker does notimpair the ability of the interfering RNA to down-regulate theexpression of target gene(s).

In addition to the silencing element(s) and any linker sequences, theinterfering RNA of the invention may comprise at least one additionalpolynucleotide sequence. In different embodiments of the invention, theadditional sequence is chosen from (i) a sequence capable of protectingthe interfering RNA against RNA processing, (ii) a sequence affectingthe stability of the interfering RNA, (iii) a sequence allowing proteinbinding, for example to facilitate uptake of the interfering RNA bycells of the insect pest species, (iv) a sequence facilitatinglarge-scale production of the interfering RNA, (v) a sequence which isan aptamer that binds to a receptor or to a molecule on the surface ofthe insect pest cells to facilitate uptake, or (v) a sequence thatcatalyses processing of the interfering RNA within the insect pest cellsand thereby enhances the efficacy of the interfering RNA. Structures forenhancing the stability of RNA molecules are well known in the art andare described further in WO2006/046148 as incorporated herein byreference.

The length of the interfering RNA of the invention needs to besufficient for uptake by the cells of an insect pest species anddown-regulation of target genes within the pest as described elsewhereherein. However, the upper limit on length may be dependent on (i) therequirement for the interfering RNA to be taken up by cells of the pestand (ii) the requirement for the interfering RNA to be processed in thecells of the pest to mediate gene silencing via the RNAi pathway. Thelength may also be dictated by the method of production and theformulation for delivery of the interfering RNA to cells. Preferably,the interfering RNA of the current invention will be between 21 and10000 nucleotides in length, preferably between 50 and 5000 nucleotidesor between 100 and 2500 nucleotides, more preferably between 80 and 2000nucleotides in length.

The interfering RNA may contain DNA bases, non-natural bases ornon-natural backbone linkages or modifications of the sugar-phosphatebackbone, for example to enhance stability during storage or enhanceresistance to degradation by nucleases. Furthermore, the interfering RNAmay be produced chemically or enzymatically by one skilled in the artthrough manual or automated reactions. Alternatively, the interferingRNA may be transcribed from a polynucleotide encoding the same. Thus,provided herein is an isolated polynucleotide encoding any of theinterfering RNAs of the current invention.

Also provided herein is an isolated polynucleotide selected from thegroup consisting of (i) a polynucleotide which comprises at least 21,preferably at least 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50,55, 60, 70, 80, 90, 100, 110, 125, 150, 175, 200, 225, 250, 300, 350,400, 450, 500, 550, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400,1500, 2000 or 3000 contiguous nucleotides of a nucleotide sequence asrepresented by any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181,189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139,5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289,298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123,132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147,126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137,185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245,152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158,159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165,167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281,200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386,387, 388, 389, or the complement thereof, or (ii) a polynucleotide whichconsists of at least 21, preferably at least 22, 23, 24, 25, 26, 27, 28,29, 30, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 110, 125, 150, 175,200, 225, 250, 300, 350, 400, 450, 500, 550, 600, 700, 800, 900, 1000,1100 or 1115 contiguous nucleotides of a nucleotide sequence asrepresented by any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181,189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139,5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289,298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123,132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147,126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184, 137,185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to 245,152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158,159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165,167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281,200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386,387, 388, 389, or the complement thereof, or (iii) a polynucleotidewhich comprises at least 21, preferably at least 22, 23 or 24, 25, 26,27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 110, 125, 150,175, 200, 225, 250, 300, 350, 400, 450, 500, 550, 600, 700, 800, 900,1000, 1100, 1200, 1300, 1400, 1500, 2000 or 3000 contiguous nucleotidesof a nucleotide sequence as represented in any of SEQ ID NOs 1, 174,404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10,43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70,23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142, 176, 182, 130, 177,183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179,210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146,125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to233, 128, 149, 184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238 to241, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253,156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163,162, 164, 266 to 269, 165, 167, 166, 270 to 273, 168, 170, 169, 274 to277, 172, 173, 278 to 281, 200, 201, 314 to 317, 186, 202, 187, 203, 306to 309, 318 to 321, 386, 387, 388, 389, or the complement thereof, sothat, when the two sequences are optimally aligned and compared, saidpolynucleotide is at least 75% preferably at least 80%, 85%, 90%, 95%,98% or 99% identical to any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175,181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31 to 34,139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12,47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17,18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25,26, 75 to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to 209, 286 to289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213, 290 to 293,123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134, 222 to 225,147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128, 149, 184,137, 185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150, 151, 242 to245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157, 254 to 257,158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164, 266 to 269,165, 167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172, 173, 278 to281, 200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309, 318 to 321,386, 387, 388, 389, or the complement thereof, or (iv) a polynucleotidewhich comprises a fragment of at least 21, preferably at least 22, 23 or24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100,110, 125, 150, 175, 200, 225, 250, 300, 350, 400, 450, 500, 550, 600,700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 2000 or 3000contiguous nucleotides of a nucleotide as represented in any of SEQ IDNOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15,204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66,21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142,176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122,144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124,133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229,127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302 to305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to 249,154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160,161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273,168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to 317,186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, or thecomplement thereof, and wherein said fragment or said complement has anucleotide sequence so that, when said fragment is optimally aligned andcompared with the corresponding fragment in any of SEQ ID NOs 1, 174,404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10,43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70,23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142, 176, 182, 130, 177,183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179,210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146,125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to233, 128, 149, 184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238 to241, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253,156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163,162, 164, 266 to 269, 165, 167, 166, 270 to 273, 168, 170, 169, 274 to277, 172, 173, 278 to 281, 200, 201, 314 to 317, 186, 202, 187, 203, 306to 309, 318 to 321, 386, 387, 388, 389, said nucleotide sequence is atleast 75% preferably at least 80%, 85%, 90%, 95%, 98% or 99% identicalto said corresponding fragment of any of SEQ ID NOs 1, 174, 404, 180,188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3,4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46,141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71to 74, 25, 26, 75 to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128,149, 184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150,151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157,254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164,266 to 269, 165, 167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172,173, 278 to 281, 200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309,318 to 321, 386, 387, 388, 389 or the complement thereof, or (v) apolynucleotide which consists of a fragment of at least 21, preferablyat least 22, 23 or 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60,70, 80, 90, 100, 110, 125, 150, 175, 200, 225, 250, 300, 350, 400, 450,500, 550, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 2000or 3000 contiguous nucleotides of a nucleotide as represented in any ofSEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to285, 294 to 297, 310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140,7, 8, 39 to 42, 9, 10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to54, 15, 204, 16, 205, 55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63to 66, 21, 22, 67 to 70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121,142, 176, 182, 130, 177, 183, 206 to 209, 286 to 289, 298 to 301, 145,122, 144, 178, 131, 179, 210 to 213, 290 to 293, 123, 132, 214 to 217,124, 133, 218 to 221, 146, 125, 134, 222 to 225, 147, 126, 135, 226 to229, 127, 148, 136, 230 to 233, 128, 149, 184, 137, 185, 234 to 237, 302to 305, 129, 138, 238 to 241, 150, 151, 242 to 245, 152, 153, 246 to249, 154, 155, 250 to 253, 156, 157, 254 to 257, 158, 159, 258 to 261,160, 161, 262 to 265, 163, 162, 164, 266 to 269, 165, 167, 166, 270 to273, 168, 170, 169, 274 to 277, 172, 173, 278 to 281, 200, 201, 314 to317, 186, 202, 187, 203, 306 to 309, 318 to 321, 386, 387, 388, 389, orthe complement thereof, and wherein said fragment or said complement hasa nucleotide sequence so that, when said fragment is optimally alignedand compared with the corresponding fragment in any of SEQ ID NOs 1,174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297,310 to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9,10, 43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205,55 to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to70, 23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142, 176, 182, 130,177, 183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131,179, 210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221,146, 125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230to 233, 128, 149, 184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238to 241, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to253, 156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265,163, 162, 164, 266 to 269, 165, 167, 166, 270 to 273, 168, 170, 169, 274to 277, 172, 173, 278 to 281, 200, 201, 314 to 317, 186, 202, 187, 203,306 to 309, 318 to 321, 386, 387, 388, 389, said nucleotide sequence isat least 75% preferably at least 80%, 85%, 90%, 95%, 98% or 99%identical to said corresponding fragment of any of SEQ ID NOs 1, 174,404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310to 313, 3, 4, 31 to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10,43 to 46, 141, 11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55to 58, 322 to 325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70,23, 24, 71 to 74, 25, 26, 75 to 78, 143, 121, 142, 176, 182, 130, 177,183, 206 to 209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179,210 to 213, 290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146,125, 134, 222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to233, 128, 149, 184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238 to241, 150, 151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253,156, 157, 254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163,162, 164, 266 to 269, 165, 167, 166, 270 to 273, 168, 170, 169, 274 to277, 172, 173, 278 to 281, 200, 201, 314 to 317, 186, 202, 187, 203, 306to 309, 318 to 321, 386, 387, 388, 389 or the complement thereof, or(vi) a polynucleotide encoding an amino acid sequence that, when the twoamino acid sequences are optimally aligned and compared, is at least 70%preferably at least 75%, 80%, 85%, 90%, 95%, 98% or 99% identical to theamino acid sequence encoded by any of SEQ ID NOs 1, 174, 404, 180, 188,2, 175, 181, 189, 27 to 30, 282 to 285, 294 to 297, 310 to 313, 3, 4, 31to 34, 139, 5, 6, 35 to 38, 140, 7, 8, 39 to 42, 9, 10, 43 to 46, 141,11, 12, 47 to 50, 13, 14, 51 to 54, 15, 204, 16, 205, 55 to 58, 322 to325, 17, 18, 59 to 62, 19, 20, 63 to 66, 21, 22, 67 to 70, 23, 24, 71 to74, 25, 26, 75 to 78, 143, 121, 142, 176, 182, 130, 177, 183, 206 to209, 286 to 289, 298 to 301, 145, 122, 144, 178, 131, 179, 210 to 213,290 to 293, 123, 132, 214 to 217, 124, 133, 218 to 221, 146, 125, 134,222 to 225, 147, 126, 135, 226 to 229, 127, 148, 136, 230 to 233, 128,149, 184, 137, 185, 234 to 237, 302 to 305, 129, 138, 238 to 241, 150,151, 242 to 245, 152, 153, 246 to 249, 154, 155, 250 to 253, 156, 157,254 to 257, 158, 159, 258 to 261, 160, 161, 262 to 265, 163, 162, 164,266 to 269, 165, 167, 166, 270 to 273, 168, 170, 169, 274 to 277, 172,173, 278 to 281, 200, 201, 314 to 317, 186, 202, 187, 203, 306 to 309,318 to 321, 386, 387, 388, 389, and wherein said polynucleotide is nolonger than 10000, 9000, 8000, 7000, 6000, 5000, 4000, 3000, 2000 or1500 nucleotides.

In preferred embodiments, the isolated polynucleotide is part of aninterfering RNA molecule, typically part of the silencing element,comprising at least one double-stranded region comprising a sense RNAstrand annealed by complementary basepairing to an antisense RNA strandwherein the sense strand of the dsRNA molecule comprises a sequence ofnucleotides complementary to a sequence of nucleotides located withinthe RNA transcript of the target gene. The sense strand of the dsRNA istherefore able to anneal to the RNA transcript and target the RNA fordegradation within the RNAi-induced silencing complex or RISC.

The polynucleotides of the invention may be inserted via routinemolecular cloning techniques into DNA constructs or vectors known in theart. Therefore, according to one embodiment, a DNA construct comprisingany of the polynucleotides of the current invention is provided.Preferably, provided herein is a DNA construct comprising apolynucleotide encoding at least one of the interfering RNAs of thecurrent invention. The DNA construct may be a recombinant DNA vector,for example a bacterial, viral or yeast vector. In a preferredembodiment of the invention, the DNA construct is an expressionconstruct and the polynucleotide is operably linked to at least oneregulatory sequence capable of driving expression of the polynucleotidesequence. The term ‘regulatory sequence’ is to be taken in a broadcontext and is intended to refer to any nucleotide sequence capable ofeffecting expression of polynucleotides to which it is operably linkedincluding but not limited to promoters, enhancers and othernaturally-occurring or synthetic transcriptional activator elements. Theregulatory sequence may be located at the 5′ or 3′ end of thepolynucleotide sequence. The term ‘operably linked’ refers to afunctional linkage between the regulatory sequence and thepolynucleotide sequence such that the regulatory sequence drivesexpression of the polynucleotide. Operably linked elements may becontiguous or non-contiguous.

Preferably, the regulatory sequence is a promoter selected from thegroup comprising but not limited to constitutive promoters, induciblepromoters, tissue-specific promoters and growth/developmentalstage-specific promoters. In one embodiment, the polynucleotide isplaced under the control of a strong constitutive promoter such as anyselected from the group comprising the CaMV35S promoter, doubled CaMV35Spromoter, ubiquitin promoter, actin promoter, rubisco promoter, GOS2promoter, Figwort mosaic virus 34S promoter.

Optionally, one or more transcription termination sequences may beincorporated in the expression construct of the invention. The term‘transcription termination sequence’ encompasses a control sequence atthe end of a transcriptional unit, which signals termination oftranscription, 3′ processing and poly-adenylation of a primarytranscript. Additional regulatory sequences including but not limited totranscriptional or translational enhancers may be incorporated in theexpression construct, for instance as with the double enhanced CaMV35Spromoter.

The present invention also encompasses a method for generating any ofthe interfering RNAs of the invention comprising the steps of (i)contacting a polynucleotide encoding said interfering RNA or a DNAconstruct comprising the same with cell-free components; or (ii)introducing (e.g. by transformation, transfection or injection) apolynucleotide encoding said interfering RNA or a DNA constructcomprising the same into a cell.

The invention thus also relates to any double stranded ribonucleotideproduced from the expression of a polynucleotide described herein.

Accordingly, also provided herein is a host cell transformed with any ofthe polynucleotides described herein. Further encompassed by the presentinvention are host cells comprising any of the interfering RNA's of thecurrent invention, any of the polynucleotides of the current inventionor a DNA construct comprising the same. The host cell may be aprokaryotic cell including but not limited to gram-positive andgram-negative bacterial cells, or an eukaryotic cell including but notlimited to yeast cells or plant cells. Preferably, said host cell is abacterial cell or a plant cell. The bacterial cell can be chosen fromthe group comprising, but not limited to, Gram positive and Gramnegative cells comprising Escherichia spp. (e.g. E. coli), Bacillus spp.(e.g. B. thuringiensis), Rhizobium spp., Lactobacillus spp., Lactococcusspp., Pseudomonas spp. and Agrobacterium spp. The polynucleotide or DNAconstruct of the invention may exist or be maintained in the host cellas an extra-chromosomal element or may be stably incorporated into thegenome of the host cell. Characteristics of particular interest inselecting a host cell for the purposes of the current invention includethe ease with which the polynucleotide or DNA construct encoding theinterfering RNA can be introduced into the host, the availability ofcompatible expression systems, the efficiency of expression, and thestability of the interfering RNA in the host.

Preferably, the interfering RNAs of the invention are expressed in aplant host cells. Preferred plants of interest include but are notlimited to cotton, potato, rice, tomato, canola, soy, sunflower,sorghum, pearl millet, corn, alfalfa, strawberries, eggplant, pepper andtobacco.

In situations wherein the interfering RNA is expressed within a hostcell and/or is used to prevent and/or control pest infestation of a hostorganism, it is preferred that the interfering RNA does not exhibitsignificant ‘off-target’ effects i.e. the interfering RNA does notaffect expression of genes within the host. Preferably, the silencingelement does not exhibit significant complementarity with nucleotidesequences other than the intended target nucleotide sequence of thetarget gene. In one embodiment of the invention, the silencing elementshows less than 30%, more preferably less than 20%, more preferably lessthan 10% and even more preferably less than 5% sequence identity withany gene of the host cell or organism. If genomic sequence data isavailable for the host organism, one can cross-check identity with thesilencing element using standard bioinformatics tools. In oneembodiment, there is no sequence identity between the silencing elementand a gene from the host cell or host organism over a region of 17, morepreferably over a region of 18 or 19 and most preferably over a regionof 20 or 21 contiguous nucleotides.

In the practical application of the invention, the interfering RNAs ofthe invention may be used for the prevention and/or control of anyinsect pest belonging to the Orders Coleoptera, Lepidoptera, Diptera,Dichyoptera, Orthoptera, Hemiptera and Siphonaptera.

Furthermore, in accordance with another aspect of the invention, thereis provided herein a composition for preventing and/or controllinginsect pest infestation comprising at least one interfering ribonucleicacid (RNA) and optionally at least one suitable carrier, excipient ordiluent, wherein the interfering RNA functions upon uptake by the pestto down-regulate the expression of a target gene within said pest. Theinterfering RNA may be any of those as disclosed elsewhere herein.Preferably, the interfering RNA comprises or consists of at least onesilencing element and said silencing element is a region ofdouble-stranded RNA comprising annealed complementary strands, onestrand of which (the sense strand) comprises a sequence of nucleotideswhich is at least partially complementary to a target nucleotidesequence within a target gene. The ‘target gene’ may be any of the pesttarget genes as disclosed elsewhere herein including but not limited togenes involved in regulating pest survival, growth, development,reproduction and pathogenicity. Alternatively, the composition comprisesat least one host cell comprising at least one interfering RNA moleculeor DNA construct encoding the same and optionally at least one suitablecarrier, excipient or diluent, wherein the interfering RNA functionsupon uptake of the host cell by the insect pest to down-regulate theexpression of a target gene within said pest.

In the practical application of the invention, the composition may beused for the prevention and/or control of any insect pest belonging tothe Orders Coleoptera, Lepidoptera, Diptera, Dichyoptera, Orthoptera,Hemiptera and Siphonaptera. The composition may therefore be in anysuitable form for application to insect pests or for application tosubstrates and/or organisms, in particular plants, susceptible toinfestation by said insect pest. In one embodiment, the composition isfor use in preventing and/or controlling pest infestation of plants orpropagation or reproductive material of plants and is thus directedtowards insect pest species that infest plants. The composition of thepresent invention is particularly effective when the insect pest belongsto the category of ‘chewing’ insects that cause considerable damage toplants by eating plant tissues such as roots, leaves, flowers, buds,twigs and the like. Examples from this large insect category includebeetles and their larvae.

The composition of the invention may be used to control insect pests atall stages of their life cycle, for example, the mature adult stage, thelarval and egg stages.

In the context of the composition of the invention, the interfering RNAmay be produced from a DNA construct, in particular an expressionconstruct as described elsewhere herein, comprising a polynucleotideencoding the same. In preferred embodiments, the interfering RNA may beproduced inside a host cell or organism engineered to express saidinterfering RNA from a polynucleotide encoding the same.

Suitable host organisms for use in the compositions of the currentinvention include but are not limited to microorganisms that are knownto colonize the environment on and/or around plants or crops of interesti.e. plants or crops susceptible to infestation by insect pest species.Such microorganisms include but are not limited to those that occupy thephylloplane (the surface of plant leaves) and/or the rhizosphere (thesoil surrounding plant roots). These microorganisms are selected so asto be capable of successfully competing with any wild-type organismspresent in the plant environment. Suitable microorganisms for use ashosts include various species of bacteria, algae and fungi. It is clearthat the chosen microorganisms must not be toxic to plants. Suchcompositions applied to plants susceptible of infestation by insect pestspecies will be ingested by the insect pests feeding on the treatedplants.

Host organisms that do not naturally colonize plants and/or theirenvironment are also within the scope of the current invention. Suchorganisms may serve only as a means to generate the interfering RNA ofthe composition. For example, in one embodiment, the interfering RNA isfermented/produced in a bacterial host and the bacteria are subsequentlyinactivated/killed. The resulting bacteria may be processed and used asan insecticidal spray in the same manner that Bacillus thuringiensisstrains have been used as an insecticide for a spray application. Incertain embodiments, a bacterial extract or lysate may be suitablypurified to leave a substantially pure interfering RNA containingextract, which is subsequently formulated into one of the compositionsof the invention. Standard extraction/purification techniques would beknown by a person skilled in the art.

Compositions of the invention may be in any suitable physical form forapplication to insects. For example, the composition may be in solidform (powder, pellet or a bait), liquid form (including a formadministered as a spray insecticide) or gel form. In a specificembodiment, the composition may be a coating, paste or powder that canbe applied to a substrate in order to protect said substrate frominfestation by insects. In this embodiment, the composition can be usedto protect any substrate or material that is susceptible to infestationby or damage caused by an insect.

The nature of the excipients and the physical form of the compositionmay vary depending on the nature of the substrate that it is desired totreat. For example, the composition may be a liquid that is brushed orsprayed onto or imprinted into the material or substrate to be treated,or a coating or powder that is applied to the material or substrate tobe treated.

In one embodiment, the composition is in the form of a bait. The bait isdesigned to lure the insect to come into contact with the composition.Upon coming into contact therewith, the composition is then internalisedby the insect, by ingestion for example and mediates RNAi to thus killthe insect. Said bait may comprise a food substance, such as a proteinbased food, for example fish meal. Boric acid may also be used as abait. The bait may depend on the species being targeted. An attractantmay also be used. The attractant may be a pheromone, such as a male orfemale pheremone for example. As an example, the pheromones referred toin the book “Insect Pheremones and their use in Pest Management” (Howseet al, Chapman and Hall, 1998) may be used in the invention. Theattractant acts to lure the insect to the bait, and may be targeted fora particular insect or may attract a whole range of insects. The baitmay be in any suitable form, such as a solid, paste, pellet or powderedform.

The bait may also be carried away by the insect back to the colony. Thebait may then act as a food source for other members of the colony, thusproviding an effective control of a large number of insects andpotentially an entire insect pest colony. This is an advantageassociated with use of the double stranded RNA of the invention, becausethe delayed action of the RNAi mediated effects on the pests allows thebait to be carried back to the colony, thus delivering maximal impact interms of exposure to the insects.

Additionally, compositions which come into contact with the insects mayremain on the cuticle of the insect. When cleaning, either an individualinsect cleaning itself or insects cleaning one another, the compositionsmay be ingested and can thus mediate their effects in the insect. Thisrequires that the composition is sufficiently stable such that theinterfering RNA remains intact and capable of mediating RNAi even whenexposed to external environmental conditions for a length of time, whichmay be a period of days for example.

The baits may be provided in a suitable “housing” or “trap”. Suchhousings and traps are commercially available and existing traps may beadapted to include the compositions of the invention. Any housing ortrap which may attract an insect to enter it is included within thescope of the invention. The housing or trap may be box-shaped forexample, and may be provided in pre-formed condition or may be formed offoldable cardboard for example. Suitable materials for a housing or trapinclude plastics and cardboard, particularly corrugated cardboard.Suitable dimensions for such a housing or trap are, for example, 7-15 cmwide, 15-20 cm long and 1-5 cm high. The inside surfaces of the trapsmay be lined with a sticky substance in order to restrict movement ofthe insect once inside the trap. The housing or trap may contain asuitable trough inside which can hold the bait in place. A trap isdistinguished from a housing because the insect can not readily leave atrap following entry, whereas a housing acts as a “feeding station”which provides the insect with a preferred environment in which they canfeed and feel safe from predators. Accordingly, in a further aspect theinvention provides a housing or trap for insects which contains acomposition of the invention, which may incorporate any of the featuresof the composition described herein.

In a further alternative embodiment, the composition may be provided inthe form of a spray. Thus, a human user can spray the pest directly withthe composition. The composition is then internalized by the insect,from where it can mediate RNA interference, thus controlling the insect.The spray is preferably a pressurized/aerosolized spray or a pump spray.The particles may be of suitable size such that they adhere to theinsect, for example to the exoskeleton, and may be absorbed therefrom.Particle size may be measured by known means, such as by use of aMastersizer, which is a commercially available device.

In a still further embodiment, the carrier is an electrostaticallycharged powder or particle which adheres to the insect. Suitable powdersand particles which are capable of adhering to an insect and thusdelivering the RNA constructs of the invention are described in detailin WO 94/00980 and WO 97/33472, both of which are incorporated herein byreference.

Alternatively, the carrier may comprise magnetic particles which adhereto the insect cuticle. Suitable magnetic particles which are capable ofadhering to an insect and thus delivering the RNA constructs of theinvention are described in detail in WO 00/01236, which reference isincorporated herein. In a still further embodiment, the carrier of thecomposition comprises metallic particles which are initiallyunmagnetised but which are capable of becoming magnetically polarisedwhen subjected to the electrical field provided by the insect body. Thismode of action is described in detail in WO 2004/049807 and isincorporated by reference herein.

Preferably, the composition incorporates a carrier which increases theuptake of the interfering RNA into the insect pest. Such a carrier maybe a lipid-based carrier, preferably comprising one or more of, oil-inwater emulsions, micelles, cholesterol, lipopolyamines and liposomes.Other agents which promote uptake of the constructs of the invention arewell known to those of skill in the art and include polycations,dextrans and (tris) cationic lipids, such as CS096, CS102 etc.Commercially available liposomes include LIPOFECTIN® and CELLFECTIN®etc. A number of suitable carriers are listed under the heading“Transfection promoting agent” in WO 03/004644 and each of the examplesprovided is hereby incorporated by reference.

In a further preferred embodiment, the carrier is a nucleic acidcondensing agent. Preferably, the nucleic acid condensing agentcomprises spermidine or protamine sulphate or a derivative thereof.Wherein the composition of the invention is for use in preventing and/orcontrolling pest infestation of a plant, the composition can contain anagriculturally suitable carrier. Such a carrier may be any material thatthe plant to be treated can tolerate, which does not cause undue damageto the environment or other organisms therein and, which allows theinterfering RNA to remain effective against the insect pest species. Inparticular, the compositions of the invention may be formulated fordelivery to plants in accordance with routine agricultural practicesused in the bioinsecticide industry. The composition may contain furthercomponents capable of performing other functions including but notlimited to (i) enhancement or promotion of uptake of the interfering RNAby cells of the pest and (ii) stabilization of the active components ofthe composition. Specific examples of such further components containedin the composition comprising the interfering RNA, are yeast tRNA oryeast total RNA.

The compositions may be formulated for direct application or as aconcentration of a primary composition that requires dilution prior touse. Alternatively, the composition may be supplied as kit comprisingthe interfering RNA or the host cell comprising or expressing the samein one container and the suitable diluent or carrier for the RNA or hostcell in a separate container. In the practical application of theinvention, the composition may be applied to a plant or any part of aplant at any stage of the plant's development. In one embodiment, thecomposition is applied to the aerial parts of a plant, for exampleduring cultivation of plant crops in a field. In a further embodiment,the composition is applied to the seeds of a plant either while they arein storage or once they are planted in the soil. It is generallyimportant to obtain good control of pests in the early stages of plantgrowth as this is the time when the plant can be most severely damagedby pest species.

The composition may be applied to the environment of an insect pest byvarious techniques including but not limited to spraying, atomizing,dusting, scattering, pouring, coating of seeds, seed treatment,introduction into the soil, and introduction into irrigation water. Inthe treatment of plants susceptible to pest infestation, the compositionmay be delivered to the plant or part of a plant before the appearanceof the pest (for the purposes of prevention), or once signs of pestinfestation begin to appear (for the purposes of pest control).

In a further embodiment of the invention, the compositions of theinvention may be formulated so as to contain at least one further activeagent. Thus, the composition may be provided as a “kit-of-parts”comprising the interfering RNA containing composition in one containerand one or more suitable active ingredients, for example a chemical orbiological pesticide, in a separate container. Alternatively, thecompositions may be provided as a mixture which are stable and to beused in conjunction with one another.

Suitable active ingredients which may act in a complementary manner tothe interfering RNA molecules of the present invention include, but arenot limited to the following: Chlorpyrifos, Allethrin, Resmethrin,Tetrabromoethyl, Dimethol-cyclopropane carboxylic acid (which aregenerally included in liquid compostions); and Hydramethylnon,Avermectin, Chlorpyrifos, Sulfuramid, Hydroprene, Fipronil (GABAreceptor), Isopropylphenyl methyl carbamate, Indoxacarb (PARA),Noviflumuron (Chitinsynthesis inhibitor), Imiprothrin (PARA), Abamectin(Glutamate-gated Chloride channel), Imidacloprid (Acethylcholinreceptor) (which are generally included in bait compositions).

In a preferred embodiment, the active ingredient is known to be apreferred insecticide in terms of health and environmentalconsiderations, such as for instance Hydramethylnon and Avermectin.

In a further embodiment of the invention, the composition is formulatedso as to contain at least one further agronomical agent, for example aherbicide or an additional pesticide. As used herein, a ‘secondpesticide’ or ‘additional pesticide’ refers to a pesticide other thanthe first or original interfering RNA molecule of the composition.Alternatively, the composition of the invention may be delivered incombination with at least one other agronomical agent, for a example aherbicide or a second pesticide. In one embodiment, the composition isprovided in combination with a herbicide selected from any known in theart, for instance glyphosate, imidazolinone, sulphonylurea andbromoxynil. In a further embodiment, the composition is provided incombination with at least one additional pesticide. The additionalpesticide may be selected from any pesticides known in the art and/ormay comprise an interfering ribonucleic acid that functions upon uptakeby a pest to down-regulate expression of a target gene in said pestspecies. In one embodiment, the target pest is an insect pest speciesand the interfering RNA is selected from any of the interfering RNAs asdescribed herein. In a further embodiment, the additional pesticidecomprises an interfering RNA that functions to down-regulate expressionof a known gene in any target pest species, not limited to insect pests.The original interfering RNA molecule of the composition and the secondor additional pesticide(s) may target the same insect pest species ormay be intended to target different insect pest species. For example,the original interfering RNA and the second pesticide may targetdifferent species of insect pest or may target different families orclasses of pest organisms, for example, fungi or nematodes or insects.It will be apparent to one skilled in the art how to test combinationsof interfering RNA molecules and other agronomical agents forsynergistic effects. In a preferred embodiment, the composition containsa first interfering RNA molecule described elsewhere herein and one ormore additional pesticides, each toxic to the same insect pest, whereinthe one or more additional pesticides are selected from a patatin, aBacillus thuringiensis insecticidal protein, a Xenorhabdus insecticidalprotein, a Photorhabdus insecticidal protein, a Bacillus laterosporousinsecticidal protein, a Bacillus spaericus insecticidal protein, and alignin, and wherein said Bacillus thuringiensis insecticidal protein isselected from the group consisting of a Cry1Ab, a Cry1C, a Cry2Aa, aCry3, a TIC851, a CryET70, a Cry22, a VIP, a TIC901, a TIC1201, aTIC407, a TIC417, a binary insecticidal protein selected from CryET33and CryET34, CryET80 and CryET76, TIC100 and TIC101, and PS149B1, andinsecticidal chimeras of any of the preceding insecticidal proteins.

The different components of the combinations described herein may beadministered, for example to a host organism susceptible to infestationby pest, in any order. The components may be delivered simultaneously orsequentially to the area or organism to be treated.

Also provided herein is a method for preventing and/or controlling pestinfestation, comprising contacting an insect pest species with aneffective amount of at least one interfering RNA wherein the RNAfunctions upon uptake by said pest to down-regulate expression of anessential pest target gene. The essential target gene may be any pestgene involved in the regulation of an essential biological processrequired by the pest to initiate or maintain infestation including butnot limited to survival, growth, development, reproduction andpathogenicity. In particular, the target gene may be any of the pestgenes as described elsewhere herein.

In the methods described herein to down-regulate expression of a targetgene in an insect pest species, double stranded RNA molecules comprisingat least 21 bp, one strand of which comprises or consists of a sequenceof nucleotides which is complementary to at least 21 contiguousnucleotides of any of SEQ ID NOs 1, 174, 404, 180, 188, 2, 175, 181,189, 27 to 30, 282 to 285, 294 to 297 or 310 to 313, or the complementthereof, can be used to down-regulate expression of the orthologoustarget gene in a coleopteran, hemipteran, lepidoteran or dipteran insectchosen from the group comprising but not limited to Leptinotarsa spp.(e.g. L. decemlineata (Colorado potato beetle), L. juncta (false potatobeetle), or L. texana (Texan false potato beetle)); Nilaparvata spp.(e.g. N. lugens (brown planthopper)); Lygus spp. (e.g. L. lineolaris(tarnished plant bug) or L. hesperus (western tarnished plant bug));Myzus spp. (e.g. M. persicae (green peach aphid)); Diabrotica spp. (e.g.D. virgifera virgifera (western corn rootworm), D. barberi (northerncorn rootworm), D. undecimpunctata howardi (southern corn rootworm) orD. virgifera zeae (Mexican corn rootworm). In the methods describedherein to down-regulate expression of a target gene in an insect pestspecies, double stranded RNA molecules comprising at least 21 bp, onestrand of which comprises or consists of a sequence of nucleotides whichis complementary to at least 21 contiguous nucleotides in any of SEQ IDNOs 1, 174, 404, 180, 188, 2, 175, 181, 189, 27 to 30, 282 to 285, 294to 297 or 310 to 313, or the complement thereof, can be used todown-regulate expression of the orthologous target gene in acoleopteran, hemipteran, lepidoteran or dipteran insect chosen from thegroup comprising but not limited to Leptinotarsa spp. (e.g. L.decemlineata (Colorado potato beetle), L. juncta (false potato beetle),or L. texana (Texan false potato beetle)); Nilaparvata spp. (e.g. N.lugens (brown planthopper)); Lygus spp. (e.g. L. lineolaris (tarnishedplant bug) or L. hesperus (western tarnished plant bug)); Myzus spp.(e.g. M. persicae (green peach aphid)); Diabrotica spp. (e.g. D.virgifera virgifera (western corn rootworm), D. barberi (northern cornrootworm), D. undecimpunctata howardi (southern corn rootworm) or D.virgifera zeae (Mexican corn rootworm), wherein the orthologous genesencode a protein having an amino acid sequence which is at least 85%,90%, 92%, 94%, 96%, 98%, 99% identical the amino acid sequence aspresented in any of SEQ ID NOs 79, 349, 405, 352 or 356 (when saidencoded proteins are optimally aligned).

In the methods described herein to down-regulate expression of a targetgene in an insect pest species, double stranded RNA molecules comprisingat least 21 bp, one strand of which comprises or consists of a sequenceof nucleotides which is complementary to at least 21 contiguousnucleotides in any of SEQ ID NOs 141, 11, 12, 47 to 50, or thecomplement thereof, can be used to down-regulate expression of theorthologous target gene in a coleopteran, hemipteran, lepidoteran ordipteran insect chosen from the group comprising but not limited toLeptinotarsa spp. (e.g. L. decemlineata (Colorado potato beetle), L.juncta (false potato beetle), or L. texana (Texan false potato beetle));Nilaparvata spp. (e.g. N. lugens (brown planthopper)); Lygus spp. (e.g.L. lineolaris (tarnished plant bug) or L. hesperus (western tarnishedplant bug)); Myzus spp. (e.g. M. persicae (green peach aphid));Diabrotica spp. (e.g. D. virgifera virgifera (western corn rootworm), D.barberi (northern corn rootworm), D. undecimpunctata howardi (southerncorn rootworm) or D. virgifera zeae (Mexican corn rootworm).

In the methods described herein to down-regulate expression of a targetgene in an insect pest species, double stranded RNA molecules comprisingat least 21 bp, one strand of which comprises or consists of a sequenceof nucleotides which is complementary to at least 21 contiguousnucleotides in any of SEQ ID NOs 141, 11, 12, 47 to 50, or thecomplement thereof, can be used to down-regulate expression of theorthologous target gene in a coleopteran, hemipteran, lepidoteran ordipteran insect chosen from the group comprising but not limited toLeptinotarsa spp. (e.g. L. decemlineata (Colorado potato beetle), L.juncta (false potato beetle), or L. texana (Texan false potato beetle));Nilaparvata spp. (e.g. N. lugens (brown planthopper)); Lygus spp. (e.g.L. lineolaris (tarnished plant bug) or L. hesperus (western tarnishedplant bug)); Myzus spp. (e.g. M. persicae (green peach aphid));Diabrotica spp. (e.g. D. virgifera virgifera (western corn rootworm), D.barberi (northern corn rootworm), D. undecimpunctata howardi (southerncorn rootworm) or D. virgifera zeae (Mexican corn rootworm), wherein theorthologous genes encode a protein having an amino acid sequence whichis at least 85%, 90%, 92%, 94%, 96%, 98%, 99% identical to the aminoacid sequence as presented in any of SEQ ID NOs 328 or 84 (when saidencoded proteins are optimally aligned).

In the methods described herein to down-regulate expression of a targetgene in an insect pest species, double stranded RNA molecules comprisingat least 21 bp, one strand of which comprises or consists of a sequenceof nucleotides which is complementary to at least 21 contiguousnucleotides in any of SEQ ID NOs 17, 18, 59 to 62, or the complementthereof, can be used to down-regulate expression of the orthologoustarget gene in a coleopteran, hemipteran, lepidoteran or dipteran insectchosen from the group comprising but not limited to Leptinotarsa spp.(e.g. L. decemlineata (Colorado potato beetle), L. juncta (false potatobeetle), or L. texana (Texan false potato beetle)); Nilaparvata spp.(e.g. N. lugens (brown planthopper)); Lygus spp. (e.g. L. lineolaris(tarnished plant bug) or L. hesperus (western tarnished plant bug));Myzus spp. (e.g. M. persicae (green peach aphid)); Diabrotica spp. (e.g.D. virgifera virgifera (western corn rootworm), D. barberi (northerncorn rootworm), D. undecimpunctata howardi (southern corn rootworm) orD. virgifera zeae (Mexican corn rootworm).

In the methods described herein to down-regulate expression of a targetgene in an insect pest species, double stranded RNA molecules comprisingat least 21 bp, one strand of which comprises or consists of a sequenceof nucleotides which is complementary to at least 21 contiguousnucleotides in any of SEQ ID NOs 17, 18, 59 to 62, or the complementthereof, can be used to down-regulate expression of the orthologoustarget gene in a coleopteran, hemipteran, lepidoteran or dipteran insectchosen from the group comprising but not limited to Leptinotarsa spp.(e.g. L. decemlineata (Colorado potato beetle), L. juncta (false potatobeetle), or L. texana (Texan false potato beetle)); Nilaparvata spp.(e.g. N. lugens (brown planthopper)); Lygus spp. (e.g. L. lineolaris(tarnished plant bug) or L. hesperus (western tarnished plant bug));Myzus spp. (e.g. M. persicae (green peach aphid)); Diabrotica spp. (e.g.D. virgifera virgifera (western corn rootworm), D. barberi (northerncorn rootworm), D. undecimpunctata howardi (southern corn rootworm) orD. virgifera zeae (Mexican corn rootworm), wherein the orthologous genesencode a protein having an amino acid sequence which is at least 85%,90%, 92%, 94%, 96%, 98%, 99% identical to the amino acid sequence aspresented in SEQ ID NOs 87 (when said encoded proteins are optimallyaligned). In the methods described herein to down-regulate expression ofa target gene in an insect pest species, double stranded RNA moleculescomprising at least 21 bp, one strand of which comprises or consists ofa sequence of nucleotides which is complementary to at least 21contiguous nucleotides in any of SEQ ID NOs 19, 20, 63 to 66, or thecomplement thereof, can be used to down-regulate expression of theorthologous target gene in a coleopteran, hemipteran, lepidoteran ordipteran insect chosen from the group comprising but not limited toLeptinotarsa spp. (e.g. L. decemlineata (Colorado potato beetle), L.juncta (false potato beetle), or L. texana (Texan false potato beetle));Nilaparvata spp. (e.g. N. lugens (brown planthopper)); Lygus spp. (e.g.L. lineolaris (tarnished plant bug) or L. hesperus (western tarnishedplant bug)); Myzus spp. (e.g. M. persicae (green peach aphid));Diabrotica spp. (e.g. D. virgifera virgifera (western corn rootworm), D.barberi (northern corn rootworm), D. undecimpunctata howardi (southerncorn rootworm) or D. virgifera zeae (Mexican corn rootworm).

In the methods described herein to down-regulate expression of a targetgene in an insect pest species, double stranded RNA molecules comprisingat least 21 bp, one strand of which comprises or consists of a sequenceof nucleotides which is complementary to at least 21 contiguousnucleotides in any of SEQ ID NOs 19, 20, 63 to 66, or the complementthereof, can be used to down-regulate expression of the orthologoustarget gene in a coleopteran, hemipteran, lepidoteran or dipteran insectchosen from the group comprising but not limited to Leptinotarsa spp.(e.g. L. decemlineata (Colorado potato beetle), L. juncta (false potatobeetle), or L. texana (Texan false potato beetle)); Nilaparvata spp.(e.g. N. lugens (brown planthopper)); Lygus spp. (e.g. L. lineolaris(tarnished plant bug) or L. hesperus (western tarnished plant bug));Myzus spp. (e.g. M. persicae (green peach aphid)); Diabrotica spp. (e.g.D. virgifera virgifera (western corn rootworm), D. barberi (northerncorn rootworm), D. undecimpunctata howardi (southern corn rootworm) orD. virgifera zeae (Mexican corn rootworm), wherein the orthologous genesencode a protein having an amino acid sequence which is at least 85%,90%, 92%, 94%, 96%, 98%, 99% identical to the amino acid sequence aspresented in SEQ ID NOs 88 (when said encoded proteins are optimallyaligned).

In the methods described herein to down-regulate expression of a targetgene in an insect pest species, double stranded RNA molecules comprisingat least 21 bp, one strand of which comprises or consists of a sequenceof nucleotides which is complementary to at least 21 contiguousnucleotides in any of SEQ ID NOs 165, 167, 166, 270 to 273, or thecomplement thereof, can be used to down-regulate expression of theorthologous target gene in a coleopteran, hemipteran, lepidoteran ordipteran insect chosen from the group comprising but not limited toLeptinotarsa spp. (e.g. L. decemlineata (Colorado potato beetle), L.juncta (false potato beetle), or L. texana (Texan false potato beetle));Nilaparvata spp. (e.g. N. lugens (brown planthopper)); Lygus spp. (e.g.L. lineolaris (tarnished plant bug) or L. hesperus (western tarnishedplant bug)); Myzus spp. (e.g. M. persicae (green peach aphid));Diabrotica spp. (e.g. D. virgifera virgifera (western corn rootworm), D.barberi (northern corn rootworm), D. undecimpunctata howardi (southerncorn rootworm) or D. virgifera zeae (Mexican corn rootworm). In themethods described herein to down-regulate expression of a target gene inan insect pest species, double stranded RNA molecules comprising atleast 21 bp, one strand of which comprises or consists of a sequence ofnucleotides which is complementary to at least 21 contiguous nucleotidesin any of SEQ ID NOs 165, 167, 166, 270 to 273, or the complementthereof, can be used to down-regulate expression of the orthologoustarget gene in a coleopteran, hemipteran, lepidoteran or dipteran insectchosen from the group comprising but not limited to Leptinotarsa spp.(e.g. L. decemlineata (Colorado potato beetle), L. juncta (false potatobeetle), or L. texana (Texan false potato beetle)); Nilaparvata spp.(e.g. N. lugens (brown planthopper)); Lygus spp. (e.g. L. lineolaris(tarnished plant bug) or L. hesperus (western tarnished plant bug));Myzus spp. (e.g. M. persicae (green peach aphid)); Diabrotica spp. (e.g.D. virgifera virgifera (western corn rootworm), D. barberi (northerncorn rootworm), D. undecimpunctata howardi (southern corn rootworm) orD. virgifera zeae (Mexican corn rootworm), wherein the orthologous genesencode a protein having an amino acid sequence which is at least 85%,90%, 92%, 94%, 96%, 98%, 99% identical to the amino acid sequence aspresented in any of SEQ ID NOs 347 or 348 (when said encoded proteinsare optimally aligned).

In the methods described herein to down-regulate expression of a targetgene in an insect pest species, double stranded RNA molecules comprisingat least 21 bp, one strand of which comprises or consists of a sequenceof nucleotides which is complementary to at least 21 contiguousnucleotides in any of SEQ ID NOs 143, 121, 142, 176, 182, 130, 177, 183,206 to 209, 286 to 289, 298 to 301, or the complement thereof, can beused to down-regulate expression of the orthologous target gene in acoleopteran, hemipteran, lepidoteran or dipteran insect chosen from thegroup comprising but not limited to Leptinotarsa spp. (e.g. L.decemlineata (Colorado potato beetle), L. juncta (false potato beetle),or L. texana (Texan false potato beetle)); Nilaparvata spp. (e.g. N.lugens (brown planthopper)); Lygus spp. (e.g. L. lineolaris (tarnishedplant bug) or L. hesperus (western tarnished plant bug)); Myzus spp.(e.g. M. persicae (green peach aphid)); Diabrotica spp. (e.g. D.virgifera virgifera (western corn rootworm), D. barberi (northern cornrootworm), D. undecimpunctata howardi (southern corn rootworm) or D.virgifera zeae (Mexican corn rootworm).

In the methods described herein to down-regulate expression of a targetgene in an insect pest species, double stranded RNA molecules comprisingat least 21 bp, one strand of which comprises or consists of a sequenceof nucleotides which is complementary to at least 21 contiguousnucleotides in any of SEQ ID NOs 143, 121, 142, 176, 182, 130, 177, 183,206 to 209, 286 to 289, 298 to 301, or the complement thereof, can beused to down-regulate expression of the orthologous target gene in acoleopteran, hemipteran, lepidoteran or dipteran insect chosen from thegroup comprising but not limited to Leptinotarsa spp. (e.g. L.decemlineata (Colorado potato beetle), L. juncta (false potato beetle),or L. texana (Texan false potato beetle)); Nilaparvata spp. (e.g. N.lugens (brown planthopper)); Lygus spp. (e.g. L. lineolaris (tarnishedplant bug) or L. hesperus (western tarnished plant bug)); Myzus spp.(e.g. M. persicae (green peach aphid)); Diabrotica spp. (e.g. D.virgifera virgifera (western corn rootworm), D. barberi (northern cornrootworm), D. undecimpunctata howardi (southern corn rootworm) or D.virgifera zeae (Mexican corn rootworm), wherein the orthologous genesencode a protein having an amino acid sequence which is at least 85%,90%, 92%, 94%, 96%, 98%, 99% identical to the amino acid sequence aspresented in any of SEQ ID NOs 330, 350 or 353 (when said encodedproteins are optimally aligned).

In the methods described herein to down-regulate expression of a targetgene in an insect pest species, double stranded RNA molecules comprisingat least 21 bp, one strand of which comprises or consists of a sequenceof nucleotides which is complementary to at least 21 contiguousnucleotides in any of SEQ ID NOs 145, 122, 144, 178, 131, 179, 210 to213, 290 to 293, or the complement thereof, can be used to down-regulateexpression of the orthologous target gene in a coleopteran, hemipteran,lepidoteran or dipteran insect chosen from the group comprising but notlimited to Leptinotarsa spp. (e.g. L. decemlineata (Colorado potatobeetle), L. juncta (false potato beetle), or L. texana (Texan falsepotato beetle)); Nilaparvata spp. (e.g. N. lugens (brown planthopper));Lygus spp. (e.g. L. lineolaris (tarnished plant bug) or L. hesperus(western tarnished plant bug)); Myzus spp. (e.g. M. persicae (greenpeach aphid)); Diabrotica spp. (e.g. D. virgifera virgifera (westerncorn rootworm), D. barberi (northern corn rootworm), D. undecimpunctatahowardi (southern corn rootworm) or D. virgifera zeae (Mexican cornrootworm).

In the methods described herein to down-regulate expression of a targetgene in an insect pest species, double stranded RNA molecules comprisingat least 21 bp, one strand of which comprises or consists of a sequenceof nucleotides which is complementary to at least 21 contiguousnucleotides in any of SEQ ID NOs 145, 122, 144, 178, 131, 179, 210 to213, 290 to 293, or the complement thereof, can be used to down-regulateexpression of the orthologous target gene in a coleopteran, hemipteran,lepidoteran or dipteran insect chosen from the group comprising but notlimited to Leptinotarsa spp. (e.g. L. decemlineata (Colorado potatobeetle), L. juncta (false potato beetle), or L. texana (Texan falsepotato beetle)); Nilaparvata spp. (e.g. N. lugens (brown planthopper));Lygus spp. (e.g. L. lineolaris (tarnished plant bug) or L. hesperus(western tarnished plant bug)); Myzus spp. (e.g. M. persicae (greenpeach aphid)); Diabrotica spp. (e.g. D. virgifera virgifera (westerncorn rootworm), D. barberi (northern corn rootworm), D. undecimpunctatahowardi (southern corn rootworm) or D. virgifera zeae (Mexican cornrootworm), wherein the orthologous genes encode a protein having anamino acid sequence which is at least 85%, 90%, 92%, 94%, 96%, 98%, 99%identical to the amino acid sequence as presented in any of SEQ ID NOs331 or 351 (when said encoded proteins are optimally aligned).

In the methods described herein to down-regulate expression of a targetgene in an insect pest species, double stranded RNA molecules comprisingat least 21 bp, one strand of which comprises or consists of a sequenceof nucleotides which is complementary to at least 21 contiguousnucleotides in any of SEQ ID NOs 128, 149, 184, 137, 185, 234 to 237,302 to 305, or the complement thereof, can be used to down-regulateexpression of the orthologous target gene in a coleopteran, hemipteran,lepidoteran or dipteran insect chosen from the group comprising but notlimited to Leptinotarsa spp. (e.g. L. decemlineata (Colorado potatobeetle), L. juncta (false potato beetle), or L. texana (Texan falsepotato beetle)); Nilaparvata spp. (e.g. N. lugens (brown planthopper));Lygus spp. (e.g. L. lineolaris (tarnished plant bug) or L. hesperus(western tarnished plant bug)); Myzus spp. (e.g. M. persicae (greenpeach aphid)); Diabrotica spp. (e.g. D. virgifera virgifera (westerncorn rootworm), D. barberi (northern corn rootworm), D. undecimpunctatahowardi (southern corn rootworm) or D. virgifera zeae (Mexican cornrootworm).

In the methods described herein to down-regulate expression of a targetgene in an insect pest species, double stranded RNA molecules comprisingat least 21 bp, one strand of which comprises or consists of a sequenceof nucleotides which is complementary to at least 21 contiguousnucleotides in any of SEQ ID NOs 128, 149, 184, 137, 185, 234 to 237,302 to 305, or the complement thereof, can be used to down-regulateexpression of the orthologous target gene in a coleopteran, hemipteran,lepidoteran or dipteran insect chosen from the group comprising but notlimited to Leptinotarsa spp. (e.g. L. decemlineata (Colorado potatobeetle), L. juncta (false potato beetle), or L. texana (Texan falsepotato beetle)); Nilaparvata spp. (e.g. N. lugens (brown planthopper));Lygus spp. (e.g. L. lineolaris (tarnished plant bug) or L. hesperus(western tarnished plant bug)); Myzus spp. (e.g. M. persicae (greenpeach aphid)); Diabrotica spp. (e.g. D. virgifera virgifera (westerncorn rootworm), D. barberi (northern corn rootworm), D. undecimpunctatahowardi (southern corn rootworm) or D. virgifera zeae (Mexican cornrootworm), wherein the orthologous genes encode a protein having anamino acid sequence which is at least 85%, 90%, 92%, 94%, 96%, 98%, 99%identical to the amino acid sequence as presented in any of SEQ ID NOs337 or 354 (when said encoded proteins are optimally aligned).

Furthermore, there is provided herein a method for preventing and/orcontrolling insect pest infestation in a field of crop plants, saidmethod comprising expressing in said plants an effective amount of aninterfering RNA as described herein.

Wherein the method is for the control of pest infestation, the phrase‘effective amount’ extends to the quantity or concentration ofinterfering RNA required to produce a phenotypic effect on the pest suchthat the numbers of pest organisms infesting a host organism are reducedand/or the amount of damage caused by the pest is reduced. In oneembodiment, the phenotypic effect is death of the pest and theinterfering RNA is used to achieve at least 20%, 30%, 40%, preferably atleast 50%, 60%, 70%, more preferably at least 80% or 90% pest mortalityas compared to control insect pests. In a further embodiment, thephenotypic effects include but are not limited to stunting of pestgrowth, cessation of feeding and reduced egg-laying. The total numbersof pest organisms infesting a host organism may thus be reduced by atleast 20%, 30%, 40%, preferably at least 50%, 60%, 70%, more preferablyat least 80% or 90% as compared with control pests. Alternatively, thedamage caused by the insect pest may be reduced by at least 20%, 30%,40%, preferably at least 50%, 60%, 70%, more preferably at least 80% or90% as compared with control insect pests. Hence, the method of theinvention can be used to achieve at least 20%, 30%, 40%, preferably atleast 50%, 60%, 70%, more preferably at least 80% or 90% pest control.

As used herein, the term ‘plant’ may include any reproductive orpropagation material for a plant. Reference to a plant may also includeplant cells, plant protoplasts, plant tissue cultures, plant calli,plant clumps and plant cells that are intact in plants or parts ofplants such as embryos, pollen, ovules, seeds, leaves, flowers,branches, fruit, kernels, ears, cobs, husks, stalks, roots, root tipsand the like.

Also provided herein is the use of the interfering ribonucleic acid(RNA) as described herein or the DNA construct as described herein forpreventing and/or controlling insect pest infestation, preferably insectpest infestation of plants.

The invention will be further understood with reference to the followingnon-limiting examples.

EXAMPLES Example 1 Identification of Target Genes in Insect Pest Species

1.1. Lygus hesperus Normalized cDNA Library and Preparation of dsRNAs inMultiwell Plates for the Screening Assays

Nucleic acids were isolated from Lygus hesperus nymphs of different lifestages, including freshly hatched nymphs 2, 4, 6 and 9 days old nymphsand adults. A cDNA library was prepared using the SMARTer™ PCR cDNASynthesis Kit, following the manufacturer's instructions (Clontech Cat.No 634925). The cDNA library was normalized using the Trimmer kit(Evrogen Cat No NK001) and cloned in the PCR4-TOPO vector (Invitrogen).The normalization of the clones introduced M2 adapters (Trimmer Kit,Evrogen, SEQ ID NO 92: AAGCAGTGGTATCAACGCAG), oppositely oriented ateach end of the clones. The recombinant vector constructs weretransformed into cells of Escherichia coli strain TOP10 (Invitrogen).The transformed cells were subsequently diluted and plated so as toobtain single colonies or clones. The clones were checked to ensure thatclone redundancy for the library did not exceed 5%. Single clones werepicked in liquid LB (Luria-broth) media, in 96-deep-well plates, andgrown overnight at 37° C. The plates also included positive (Lh423) andnegative (FP) control clones.

To generate the dsRNA, sense and antisense DNA fragments, containing T7promoter sequence, were generated by PCR. In brief, per clone, 1 μl ofbacterial suspension was dispensed in multiwell PCR plates containingREDTaq® (Sigma Cat No D4309) and primers oGCC2738 (SEQ ID NO 93:AAGCAGTGGTATCAACGCAG) and oGCC2739 (SEQ ID NO 94:GCGTAATACGACTCACTATAGGAAGCAGTGGTATCAACGCAG) based on the M2 and theT7-M2 sequences respectively. The PCR reaction was followed by in vitrotranscription, where per clone, 6 μl PCR product were added to 9 μlRiboMAX™ Large Scale RNA Production System-T7 (Promega Cat No P1300) andincubated overnight at 37° C. The final dsRNA solution was diluted 2times in L. hesperus sucrose diet, containing 15% sucrose and 5 μg/μlyeast tRNA (Invitrogen Cat No 15401-029) and used for screening. ThedsRNA corresponding to the positive Lh423 control clone is SEQ ID NO 101and to the negative FP control clone is SEQ ID NO 104 (see Table 4).

1.2. Screen for Novel and Potent Lygus hesperus Target Genes Using adsRNA Expression cDNA Library

A new screening assay for potent Lygus hesperus targets has beendeveloped. The assay set-up was as follows: each well of a 96-well platehouses a one-day-old L. hesperus nymph exposed to a parafilm sachetcontaining sucrose diet which includes either test dsRNA or controldsRNA in the presence of tRNA. Each plate contained dsRNA from 90different clones, 3×Lh423 (positive control) and 3×FP (fluorescentprotein; negative control). Each clone (test dsRNA) was replicated overthree plates. After three days exposure, the nymphal survival number wasrecorded and the diet replaced with fresh rearing (complex) diet inabsence of dsRNA. The mortality was assessed at days 4, 6 and 8. Anidentical set up was used for the first and second round confirmationassays, with 8 and 20 insects respectively, with one nymph per well.

The assay system was validated using dsRNA corresponding to Lh423 targetas the positive control and a fluorescent protein dsRNA as the negativecontrol: over 90% were true positives and under 5% were false positives,respectively.

Twenty 96 well-plates, named Lh001 to Lh020 (see bottom line in FIGS. 1& 2), containing 1800 individual clones have been tested. 205 candidateswere identified and tested in a first confirmation assay. Setting thethreshold at showing ≧50% mortality, 41 independent clones wereidentified and progressed to a second round of confirmation. In theassay, the clones were compared to the positive controls Lh423 (RpL19)and Lh105.2 (Sec23) and the negative control Pt (encoding a coralfluorescent protein). The dsRNA corresponding to the positive (Lh423)control clone is SEQ ID NO 101, to the positive Lh105.2 control clone isSEQ ID NO 102 and to the negative (Pt) control clone is SEQ ID NO 104(see Table 4).

Second round confirmation assays, testing 20 insects/test dsRNA, wereinitiated for all the test dsRNAs displaying ≧50% mortality in the firstconfirmation (FIGS. 1 and 2). Candidate targets corresponding to theconfirmed test dsRNAs were named with an “Lhxxx number” (see Table 1).Using the same cut-off at ≧50% mortality, 15 targets were confirmed inthe first screen.

A second screen for identifying more Lygus hesperus targets wasperformed. The results of the second round confirmation assays arerepresented in FIG. 12. Using the same cut-off at 50% mortality, severaltargets were confirmed in the second screen (see Table 1 C).

1.3. Identification of Lygus Targets

In parallel to the confirmation insect assays, the inserts correspondingto the positive clones were sequenced and BlastX searches against bothDrosophila and Tribolium protein databases were used to confirm theidentity of the targets. Table 1 provides a summary of thebio-informatics analysis and current annotation of the novel identifiedL. hesperus target sequences.

Fifteen novel L. hesperus targets were identified in the first screenand 11 novel L. Hesperus targets were identified in the second screen.All targets exhibit high potency against L. hesperus nymphs indicatingthat the cDNAs encoding double-stranded RNAs contained therein areessential for pest survival and thus represent target genes of interestfor the purposes of pest control. The DNA sequences and deduced aminoacid sequences of these target genes were therefore determined and areprovided in Tables 2 and 3 respectively.

Lh594, the Lygus hesperus orthologue of Drosophila troponin I, involvedin muscle contraction—and therefore absent in plants—, represents anovel class of target belonging to an animal specific physiologicalpathway not yet explored for GM-RNAi. In the fruit fly, troponin I isdescribed as a haplo-insufficient gene, displaying a mutant phenotype inthe heterozygote state. Such genes may be particularly susceptible toreduced mRNA expression levels and as such can be considered as idealRNAi targets.

In this Lh594 pathway, eight targets were selected (see Table 1B). Foreach target, up to 4 pairs of degenerated PCR primers were designedbased on the alignments of the sequences of various insects, includingbee, Tribolium and aphid. The primers are being used to amplifyfragments from Lygus hesperus targets. The DNA sequences and deducedamino acid sequences of these target genes were determined and areprovided in Tables 2 and 3 respectively.

TABLE 1 Lygus hesperus novel targets ranked in % mortality according tothe second confirmation assay results (first screen). rank 2nd Target IDconfirmation Best Drosophila hit NAME SYMBOL Lh594 1 CG7178 wings up A(troponin I) wupA Lh618 2 CG2168 ribosomal protein S3A RpS3A Lh609 3CG4087 ribosomal protein LP1 RpLP1 Lh595 4 — no Drosophila hit found,Lygus specific target/sequence Lh611 5 CG6779 ribosomal protein S3 RpS3Lh560 6 CG10423 ribosomal protein S27 RpS27 Lh596 7 — no Drosophila hitfound, Lygus RpL34b specific target/sequence Lh615 8 CG11522 ribosomalprotein L6 RpL6 Lh617 9 CG7283 ribosomal protein L10Ab RpL10Ab Lh612 10CG13389 ribosomal protein S13 RpS13 Lh246 11 CG3195 ribosomal proteinL12 RpL12 Lh429 12 CG8900 ribosomal protein S18 RpS18 Lh610 13 CG5502ribosomal protein L4 RpL4 Lh597 14 no hit found Lh598 15 CG34069mitochondrial cytochrome c mt:Coll oxidase subunit II Lh614 — CG7610 ATPsynthase-γ chain ATPsyn-γ

TABLE 1B Lygus hesperus novel targets in Lh594 pathway Target ID BestDrosophila hit(s) NAME SYMBOL Lh619 CG7107 troponin T (upheld) up Lh620CG17927 myosin heavy chain Mhc Lh621 CG4843 tropomyosin2 (Tm2) Tm2 Lh622CG3201 myosin light chain Mlc-c cytoplasmic Lh623 CG3595 spaghettisquash sqh Lh624 CG15792 zipper zip Lh625 *CG2981, CG7930, troponin CCG9073, CG6514, CG12408 Lh626 *CG9073, CG7930, troponin C CG2981,CG12408, CG6514 *unclear: multiple hits in family - ranked accordinge-value

TABLE 1C Lygus hesperus novel targets ranked in % mortality according tothe second confirmation assay results (second screen). Best Target rank2nd Drosophila ID confirmation hit NAME SYMBOL Lh631 1 CG6846 Ribosomalprotein L26 RpL26 Lh634.2 2 CG12775 Ribosomal protein L21 RpL21 Lh634.13 CG12775 Ribosomal protein L21 RpL21 Lh630 4 CG11271 Ribosomal proteinS12 RpS12 Lh632 5 CG2998 Ribosomal protein RpS28b S28b Lh618.2 6 CG2168Ribosomal protein RpS3A S3A Lh629 7 CG4651 Ribosomal protein L13 RpL13Lh633.2 8 CG17521 Ribosomal protein L10 RpL10 Lh628 9 CG17489 Ribosomalprotein L5 RpL5 Lh633 10 CG17521 Ribosomal protein L10 RpL10 Lh627 11CG2033 Ribosomal protein RpS15A S15Aa1.4. Full Length cDNA Cloning by RACE (Rapid Amplification of cDNA Ends)

In order to clone full length cDNA, starting from a known clone ofinternal fragment from the most potent targets, the 5′/3′ RACE kit wasused (Roche, Cat. No. 1 734 792; based on Sambrook, J. & Russell, D. M).The standard protocol, described in the Instruction Manual, wasfollowed. Briefly, for a 5′ RACE, a target sequence specific antisenseprimer was designed on the known sequence and used for a first strandcDNA synthesis, using Lygus RNA as template. A tail was added to thefirst strand cDNA and used as an anchor for the second strand synthesisand amplification of an unknown end portion of the transcript. For a 3′RACE, an oligo dT anchor primer was used for the first strand cDNAsynthesis. For the 5′ and 3′ RACEs, nested primers, specific to thetarget sequence were used in a second PCR reaction. The PCR fragmentswere analysed on agarose gel, purified, cloned and sequenced forconfirmation.

Full length cDNA sequences corresponding to the targets were assembledin VectorNTi, a fully integrated sequence analysis software package forDNA sequence analysis (Invitrogen).

Example 2 In Vitro Production of Double-Stranded RNAs for Gene Silencing

2.2. Production of dsRNAs Corresponding to the Partial Sequences of theLygus hesperus Target Genes

Double-stranded RNA was synthesized in milligram quantities. First, twoseparate 5′ T7 RNA polymerase promoter templates (a sense template andan antisense template) were generated by PCR. PCRs were designed andcarried out so as to produce sense and antisense templatepolynucleotides, each having the T7 promoter in a different orientationrelative to the target sequence to be transcribed.

For each of the target genes, the sense template was generated using atarget-specific T7 forward primer and a target-specific reverse primer.The antisense templates were generated using target-specific forwardprimers and target-specific T7 reverse primers. The sequences of therespective primers for amplifying the sense and antisense templates viaPCR for each of the target genes are provided in Table 4. The PCRproducts were analysed by agarose gel electrophoresis and purified. Theresultant T7 sense and antisense templates were mixed and transcribed bythe addition of T7 RNA polymerase. The single-stranded RNAs produced bytranscription from the templates were allowed to anneal, were treatedwith DNase and RNase, and were purified by precipitation. The sensestrand of the resulting dsRNA produced from each of the target genes isprovided in Table 4.

2.2. Survival Analysis Assays for Novel Lygus hesperus Targets

To enable ranking according to potency, in vitro dsRNAs corresponding tothe novel targets were synthesized and applied to L. hesperus in 10 dayssurvival analysis bioassays. Briefly, one day old L. hesperus nymphswere placed in 96 well-plates with sucrose seals containing 0.5 μg/μltarget dsRNA, supplemented with 5 μg/μl yeast tRNA. The plates wereincubated for 3 days under standard Lygus rearing conditions. At day 3,6 and 8, the diet seals were refreshed with seals containing Lygus dietonly. Lh423 (RpL19) was used as positive control and GFP dsRNA andsucrose diet were used as negative controls.

The results from the survival analyses confirmed the data from the firstand second confirmation assays. Lh594 was established as a highly potenttarget, with activity and speed-to-kill stronger than the strong controlLh423.

So far, the Lygus screen for novel targets identified new targets withactivities higher or in the range of the positive control Lh423, theseinclude Lh429, Lh594, Lh609, Lh610, Lh611, Lh617 and Lh618. Themortality induced by these targets is show in the FIGS. 3 and 4.

To allow a more precise ranking of the targets according to theiractivity, dose response concentration analyses were made. The noveltargets were tested in in vitro assays, with concentrations ranging from0.4 to 0.025 μg/μl. Per condition, 24 one day old nymphs were tested inthe 96 well-plate set-up, in sucrose diet supplemented with dsRNA andtRNA carrier. The results are presented as % survival over a 10 dayexperiment (FIGS. 5 to 9) and are summarized in Table 5.

Based on the concentration curve analyses, the targets were ranked bycomparison to the bench mark controls Lh423 and Lh105 (Table 5).

TABLE 5 Lygus novel targets ranking according to DRCs and compared tobench mark targets Lh423 & Lh105. Potency expressed as μg/μl TargetdsRNA needed to reach 90% kill at ID day 7 Lh594 0.025 (at day 6) Lh6180.05-0.1 Lh612 0.05 Lh615 0.05 Lh423 0.1 Lh595 0.1 Lh560 0.1 Lh610 0.1Lh617 0.1 Lh105 0.2 Lh614 0.2 (at day 6)  Lh611 0.2 Lh596 0.3 Lh609 NDLh429 ND

The potency of Lh594 was further confirmed. This target effect isclearly observed at least one day before the other targets and the benchmark positive control Lh105 and Lh423. Because Lh594 was highly potent,the LD50 was not reached in the standard DRC experiment, withconcentration ranging from 0.4 to 0.025 μg/μl dsRNA (FIG. 6), the Lh594experiment was therefore repeated, including lower concentrationsranging from 0.05 to 0.001 μg/μl dsRNA (FIG. 10). In conclusion, Lh594activity was observed at concentration as low as 0.0025 μg/μl and about90% kill (corresponding to about 10% survival) was obtained at day 6with 0.025 μg dsRNA.

To further explore the potency of Lh594 and the role of tRNA carrier inthe RNAi response in Lygus hesperus, additional in vitro feeding assayswere set up in the absence of carrier tRNA. Lh594, Lh423 (bench markcontrol) and GFP (negative control) dsRNAs were produced in vitro, usingthe standard method. The dsRNAs were purified and tested at 5 μg/μl inthe absence of tRNA (FIG. 11 A).

In absence of tRNA, targets Lh594 and Lh423, induced high lethality inLygus nymphs. The results from this experiment have been sincereproduced. Target dsRNA was able to induce RNAi-by-feeding effects inLygus nymphs in the absence of tRNA.

To investigate the activity of dsRNA at lower concentrations in theabsence of carrier tRNA, additional experiments were set up, usingdecreasing amounts of dsRNA (FIG. 11 B).

A similar approach was followed for the Lygus targets that wereidentified in the second screen. To allow a ranking of the targetsaccording to their activity, dose response concentration analyses weremade. The novel targets were tested in in vitro assays, withconcentrations ranging from 0.5 to 0.05 μg/μl. Per condition, 24 one dayold nymphs were tested in the 96 well-plate set-up, in sucrose dietsupplemented with dsRNA and tRNA carrier. The results are presented as %survival over a 9 day experiment (FIGS. 15 A-D). Lh594 and Lh423 havebeen included in the assay as a reference targets. The results aresummarized in Table 6. Based on the concentration curve analyses, thetargets were ranked by comparison to the bench mark control Lh423.

TABLE 6 Lygus novel targets from second screen-ranking according to DRCsand compared to bench mark targets Lh423 & Lh594. Target Potencyexpressed as μg/μl dsRNA ID needed to reach 90% kill at day 7 Lh5940.025 (at day 6) Lh634 0.1 Lh423 0.1 Lh631 0.4 Lh633 0.4 Lh627 0.5 Lh6280.5 Lh630 0.5 Lh632 0.5 Lh629 ND

Example 3 Troponin Pathway Screen

To enable testing of the Troponin pathway targets, in vitro produceddsRNAs corresponding to Lh619, Lh620, Lh621, Lh622, Lh622, Lh623, Lh624,Lh625 and Lh626 were synthesized and applied to L. hesperus in 10 dayssurvival analysis bioassays. Briefly, one day old L. hesperus nymphswere placed in 96 well-plates with sucrose seals containing 0.5 μg/μltarget dsRNA, supplemented with 5 μg/μl yeast tRNA. The plates wereincubated for 3 days under standard Lygus rearing conditions. At day 3,6 and 8, the diet seals were refreshed with seals containing Lygus dietonly. Lh594 (Troponin I) was used as positive control and GFP dsRNA andsucrose diet were used as negative controls (FIG. 13). Four targets werethen included in dose response curve analyses in an in vitro assay, withconcentrations ranging from 0.4 to 0.025 μg/μl. Per condition, 24 oneday old nymphs were tested in the 96 well-plate set-up, in sucrose dietsupplemented with dsRNA and tRNA carrier. The results are presented as %survival over a 10 day experiment (FIGS. 14 A-B).

Example 4 Identification of Target Genes in Leptinotarsa decemlineata

4.1. Leptinotarsa decemlineata Normalized cDNA Library and Preparationof dsRNAs in Multiwell Plates for the Screening Assays

Nucleic acids were isolated from Leptinotarsa decemlineata larvae ofdifferent stages. A cDNA library was prepared using the SMARTer™ PCRcDNA Synthesis Kit, following the manufacturer's instructions (ClontechCat. No 634925). The cDNA library was normalized using the Trimmer kit(Evrogen Cat No NK001) and cloned in the PCR®-BLUNTII-TOPO® vector(Invitrogen). The normalization of the clones introduced M2 adapters(Trimmer Kit, Evrogen, SEQ ID NO 92: AAGCAGTGGTATCAACGCAG), oppositelyoriented at each end of the clones. The recombinant vector constructswere transformed into cells of Escherichia coli strain TOP10(Invitrogen). The transformed cells were subsequently diluted and platedso as to obtain single colonies or clones. The clones were checked toensure that clone redundancy for the library did not exceed 5%. Singleclones were inoculated into liquid LB (Luria-broth) media, in 96-wellplates, and grown overnight at 37° C. The plates also included positive(Ld513) and negative (FP) control clones.

To generate the dsRNA, sense and antisense DNA fragments, containing T7promoter sequence, were generated by PCR. In brief, per clone, 1 μl ofbacterial suspension was dispensed in multiwell PCR plates containingREDTaq® (Sigma Cat No D4309) and primers oGCC2738 (SEQ ID NO 93:AAGCAGTGGTATCAACGCAG) and oGCC2739 (SEQ ID NO 94:GCGTAATACGACTCACTATAGGAAGCAGTGGTATCAACGCAG) based on the M2 and theT7-M2 sequences, respectively. The PCR reaction was followed by in vitrotranscription, where, per clone, 6 μl PCR product was used in a 20 μlreaction volume containing the transcription reagents provided by theRiboMAX™ Large Scale RNA Production System-T7 kit (Promega Cat No P1300)and incubated overnight at 37° C. The final dsRNA solution was dilutedin sterile Milli-Q water and used for screening. The dsRNA correspondingto the positive Ld513 control clone is SEQ ID NO 400 (see Table 9) andto the negative FP control clone is SEQ ID NO 104 (see Table 4).

4.2. Screen for Novel and Potent Leptinotarsa decemlineata Target GenesUsing a dsRNA Expression cDNA Library

Each well of a 48-well plate contained 0.5 mL artificial diet pretreatedwith a topical overlay of 25 μl (or 1 μg) of the test or control dsRNA.One L2 larva was placed in each well and 3 larvae were tested per clone.CPB survival numbers were assessed at days 4, 7 and 10.

In a second bioassay, CPB larvae were fed on diet treated with topicallyapplied test dsRNA generated from clones derived from a normalized cDNAlibrary. One larva was placed in a well of a 48-well multiplatecontaining 0.5 mL diet pretreated with a topical overlay of 25 μL of a40 ng/μL dsRNA solution. A total of twenty-four larvae were tested pertreatment (clone). The number of surviving insects were assessed at days4, 5, 6, 7, 8 & 11. The larval mortality percentage was calculatedrelative to day 0 (start of assay) (see FIG. 21).

4.3. Identification of L. decemlineata Beetle Targets

The new target sequences from the screen in 5.2. and the targetsequences corresponding to the troponin pathway targets, orthologuous tothe Lygus Lh594, Lh619 and Lh620 sequences, have been identified in L.decemlineata. The primers which provided relevant cDNA fragment forLd594 are listed in Table 17. The cDNA sequences and deduced amino acidsequences of these target genes were determined and are provided inTables 7 and 9 respectively.

4.4. Production of dsRNAs Corresponding to the Partial Sequences of theL. decemlineata Target Genes

dsRNA was synthesized using the primers as provided in Table 9. Thesense strand of the resulting dsRNA produced from the target genes isprovided in Table 9.

4.5. Survival Analysis Assays for Novel L. decemlineata Targets

Early Larval Assay

Synthetic dsRNAs were produced for the 3 targets, Ld594, Ld619 andLd620, and were tested in a feeding assay on CPB larvae (see FIG. 16). A10 day assay was performed in 48 well plates, on artificial diet (basedon Gelman et al, J Ins Sc, 1:7, 1-10: Artificial diets for rearing theColorado Potato Beetle), supplemented with 1 μg dsRNA/well, with 12larvae per condition.

A clear effect on the development of the larvae could be observed. Asecond assay was set up to investigate the effect of these dsRNAs duringthe course of pupation and metamorphosis (see pupation assayunderneath).

Pupation Assay

A CPB pupation assay was set up to investigate the effect of RNAiknock-down of Ld594, Ld619 and Ld620 during pupation and metamorphosis.Fourth instar larvae were fed 1 μg in vitro synthesized dsRNA dispensedon a potato leaf disk and were then transferred to a box containinguntreated fresh potato leaves. Four days later the surviving insectswere placed on vermiculite to allow pupation. Lh594 treated insects wereslow, smaller and mostly were unable to go through pupation. Thehatching of the pupa was assessed at the end of the experiment. For theuntreated control 24 larvae pupated and all hatched into healthy adults.For Ld620, a decrease in numbers of larvae progressing into pupation wasobserved. For the three targets tested, no larvae progressed intohealthy pupae and none emerged into adult. Dead insects recovered fromthe vermiculite showed various degrees of malformations (FIG. 17).

Ld594, Ld619 and Ld620, first appeared as not lethal targets in the CPBlarval assay, although a reduction of vitality was clearly observed inthe dsRNA treated insects. On the other hand, in the pupation assay, all3 targets induced strong effects and inhibited the entry in pupationand/or metamorphosis.

Adult Assay

To assess activity of Ld594, Ld619 and Ld620 in CPB adults, a leaf discassay was set up. A potato leaf disc (1.7 cm diameter) was painted withdsRNA or controls and was placed in a 3.5 cm Petri dish with one adultbeetle. The next day a fresh treated leaf disc was provided to theinsects. On the third day, the adults were transferred to a boxcontaining enough fresh, untreated potato leaves to sustain the survivalof the untreated controls. Per treatment, 6 adults were tested and thenumbers of survivors and moribund insects were counted at regularintervals from day 6 to day 13. The insects were considered moribund ifthey were unable to right themselves after being placed on their back.Despite the relatively high level of background in the negative controlin this particular assay, clear effects were observed for the insectsthat had been exposed to Ld594 or Ld619 dsRNAs (FIG. 18).

Example 5 Identification of Target Genes in Nilaparvata lugens

5.1 Identification of Nilaparvata lugens Targets

New target sequences, corresponding to Troponin pathway targets andnamed NI594 (Troponin I), NI619 (Troponin T) and NI626 (Troponin C) havebeen identified in brown plant hopper, Nilaparvata lugens. Orthologoussequences of the Lygus genes, named NI594 (Troponin I), NI619 (TroponinT) and NI625/626 (Troponin C), were cloned through degenerated primerPCR, using BPH cDNA as template. In addition, full length cDNA wasidentified for NI594, using RACE (see above for method). AmpliTaq GoldPCR system (Applied Biosystems) was used following the manufactures'instructions and with standard conditions for the degenerate primer PCRreactions, typically as follows: 1 cycle with 10 minutes at 95° C.,followed by 40 cycles with 30 seconds at 95° C., 1 minute at 50° C. and1 minute at 72° C., followed by 10 minutes at 72° C. To increase therate of success, up to 10 different degenerated primers, forward andreverse, were designed, based on alignments of orthologous sequences inother species, and used in various combinations. PCR fragments obtainedwere purified from the gel by gel extraction kit (Qiagen Cat. No 28706)and cloned into a TOPO TA vector (Invitrogen). The clones were sequencedand the consensus sequences were used in Blast searches against variousavailable insect sequence databases to confirm the relevance of theinsert. The degenerated primers that resulted in successfulamplification are listed in Table 18.

The DNA sequences and deduced amino acid sequences of these target genesand one other target gene (NI537) were determined and are provided inTables 10 and 11 respectively.

5.2 Production of dsRNAs Corresponding to the Partial Sequences of theNilaparvata lugens Target Genes

dsRNA was synthesized using the primers as provided in Table 12. Thesense strand of the resulting dsRNA produced from each of the targetgenes is provided in Table 12.

5.3 Survival Analysis Assays for Novel Nilaparvata lugens Targets

dsRNAs were synthesized and tested in the previously optimized BPHRNAi-by-feeding assays, in the presence of the zwitterionic detergent,CHAPSO, at 0.1% final concentration. The dsRNAs were tested at 0.5 μg/μlfinal concentration. NI537, a potent target in the BPH assays was usedas bench mark target in the assay. The insect survival was assessed overthe course of 9 days.

The results of the bioassay showed that in BPH NI1594, NI619 and NI626were also potent RNAi targets in BPH (FIG. 19).

Example 6 Identification of Target Genes in Acyrthosiphon pisum

6.1 Identification of Acyrthosiphon pisum Targets

New target sequences have been identified in aphids and were namedAp423, Ap537, Ap560 and Ap594, following the same nomenclature: “Apxxx”,where “Ap” corresponds to Acyrthosiphon pisum and “xxx” to the ID of thetarget. Primers were designed based on public domain gene prediction inAphidBase, an on-line resource (Table 13).

The DNA sequences and deduced amino acid sequences of these target geneswere determined and are provided in Tables 14 and 15 respectively.

6.2 Production of dsRNAs Corresponding to the Partial Sequences of theAphid Target Genes

dsRNA was synthesized using the primers as provided in Table 16. Thesense strand of the resulting dsRNA produced from each of the targetgenes is provided in Table 16.

6.3 Survival Analysis Assays for Novel Aphid Targets

RNAi-by-feeding was tested in Acyrthosiphon pisum (pea aphid) with 4targets Ap594, Ap423, Ap560, Ap537. The sequences were amplified by PCRusing primers, designed on public domain sequence information found inAphidBase, an on-line resource, and cDNA prepared from aphids. Thesynthetic dsRNAs were prepared and tested at a final concentration of0.5 μg/μl in presence of 5 ng/μ1 yeast tRNA in a sucrose diet. Tenneonate pea aphid nymphs were placed in a small Petri dish (32 mm).Fifty μl diet (with tRNA and dsRNA) was pipetted on top of the firstlayer of parafilm. A second layer of parafilm covered the diet andcreated a feeding sachet where the aphids could feed. Per target fivereplicates of 10 neonate nymphs were set-up. GFP dsRNA was used as anegative control. The diet was refreshed on day 4 and 7 of the assaysand survival was assessed (FIG. 20).

TABLE 2 Target ID cDNA Sequence (sense strand) 5′ → 3′ Lh594 SEQ ID NO 1Lh609 SEQ ID NO 3 Lh610 SEQ ID NO 5 Lh610 (b) SEQ ID NO 139 Lh611 SEQ IDNO 7 Lh611 (b) SEQ ID NO 140 Lh617 SEQ ID NO 9 Lh618 SEQ ID NO 11 Lh618(b) SEQ ID NO 141 Lh429 SEQ ID NO 13 Lh423 SEQ ID NO 95 Lh105.2 SEQ IDNO 96 Lh560 SEQ ID NO 15 Lh615 SEQ ID NO 17 Lh612 SEQ ID NO 19 Lh246 SEQID NO 21 Lh597 SEQ ID NO 23 Lh598 SEQ ID NO 25 Lh619 SEQ ID NO 121 Lh619(b) SEQ ID NO 142 Lh619 (c) SEQ ID NO 143 Lh620 SEQ ID NO 122 Lh620 (b)SEQ ID NO 144 Lh620 (c) SEQ ID NO 145 Lh621 SEQ ID NO 123 Lh622 SEQ IDNO 124 Lh623 SEQ ID NO 125 Lh623 (b) SEQ ID NO 146 Lh624 SEQ ID NO 126Lh624 (b) SEQ ID NO 147 Lh625 SEQ ID NO 127 Lh625 (b) SEQ ID NO 148Lh626 SEQ ID NO 128 Lh626 (b) SEQ ID NO 149 Lh614 SEQ ID NO 129 Lh627SEQ ID NO 150 Lh628 SEQ ID NO 152 Lh629 SEQ ID NO 154 Lh630 SEQ ID NO156 Lh631 SEQ ID NO 158 Lh632 SEQ ID NO 160 Lh633.1 SEQ ID NO 162Lh633.2 SEQ ID NO 163 Lh634.1 SEQ ID NO 165 Lh634.2 SEQ ID NO 167Lh595.1 SEQ ID NO 168 Lh595.2 SEQ ID NO 170 Lh596 SEQ ID NO 172

TABLE 3 Corresponding amino acid sequence of cDNA Target ID clone asrepresented in Table 2 Lh594 SEQ ID NO 79 Lh609 SEQ ID NO 80 Lh610 SEQID NO 81 Lh610 (b) SEQ ID NO 326 Lh611 SEQ ID NO 82 Lh611 (b) SEQ ID NO327 Lh617 SEQ ID NO 83 Lh618 SEQ ID NO 84 Lh618 (b) SEQ ID NO 328 Lh429SEQ ID NO 85 Lh429 (b) SEQ ID NO 329 Lh423 SEQ ID NO 99 Lh105.2 SEQ IDNO 100 Lh560 SEQ ID NO 86 Lh615 SEQ ID NO 87 Lh612 SEQ ID NO 88 Lh246SEQ ID NO 89 Lh597 SEQ ID NO 90 Lh598 SEQ ID NO 91 Lh619 SEQ ID NO 330Lh620 SEQ ID NO 331 Lh621 SEQ ID NO 332 Lh622 SEQ ID NO 333 Lh623 SEQ IDNO 334 Lh624 SEQ ID NO 335 Lh625 SEQ ID NO 336 Lh626 SEQ ID NO 337 Lh614SEQ ID NO 338 Lh627 SEQ ID NO 339 Lh628 SEQ ID NO 340 Lh629 SEQ ID NO341 Lh630 SEQ ID NO 342 Lh631 SEQ ID NO 343 Lh632 SEQ ID NO 344 Lh633.1SEQ ID NO 345 Lh633.2 SEQ ID NO 346 Lh634.1 SEQ ID NO 347 Lh634.2 SEQ IDNO 348

TABLE 4 dsRNA: sense strand represented Primers by equivalent PrimersForward Reverse DNA Sequence Target ID 5′ → 3′ 5′ → 3′ 5′ → 3′ Lh594 SEQID NO 27 SEQ ID NO 28 SEQ ID NO 2 SEQ ID NO 29 SEQ ID NO 30 Lh609 SEQ IDNO 31 SEQ ID NO 32 SEQ ID NO 4 SEQ ID NO 33 SEQ ID NO 34 Lh610 SEQ ID NO35 SEQ ID NO 36 SEQ ID NO 6 SEQ ID NO 37 SEQ ID NO 38 Lh611 SEQ ID NO 39SEQ ID NO 40 SEQ ID NO 8 SEQ ID NO 41 SEQ ID NO 42 Lh617 SEQ ID NO 43SEQ ID NO 44 SEQ ID NO 10 SEQ ID NO 45 SEQ ID NO 46 Lh618 SEQ ID NO 47SEQ ID NO 48 SEQ ID NO 12 SEQ ID NO 49 SEQ ID NO 50 Lh429 SEQ ID NO 51SEQ ID NO 52 SEQ ID NO 14 SEQ ID NO 53 SEQ ID NO 54 Lh423 SEQ ID NO 105SEQ ID NO 106 SEQ ID NO 101 SEQ ID NO 107 SEQ ID NO 108 Lh105.2 SEQ IDNO 109 SEQ ID NO 110 SEQ ID NO 102 SEQ ID NO 111 SEQ ID NO 112 GFP SEQID NO 113 SEQ ID NO 114 SEQ ID NO 103 SEQ ID NO 115 SEQ ID NO 116 Pt SEQID NO 117 SEQ ID NO 118 SEQ ID NO 104 SEQ ID NO 119 SEQ ID NO 120 Lh560SEQ ID NO 55 SEQ ID NO 56 SEQ ID NO 16 SEQ ID NO 57 SEQ ID NO 58 Lh615SEQ ID NO 59 SEQ ID NO 60 SEQ ID NO 18 SEQ ID NO 61 SEQ ID NO 62 Lh612SEQ ID NO 63 SEQ ID NO 64 SEQ ID NO 20 SEQ ID NO 65 SEQ ID NO 66 Lh246SEQ ID NO 67 SEQ ID NO 68 SEQ ID NO 22 SEQ ID NO 69 SEQ ID NO 70 Lh597SEQ ID NO 71 SEQ ID NO 72 SEQ ID NO 24 SEQ ID NO 73 SEQ ID NO 74 Lh598SEQ ID NO 75 SEQ ID NO 76 SEQ ID NO 26 SEQ ID NO 77 SEQ ID NO 78 Lh619SEQ ID NO 206 SEQ ID NO 207 SEQ ID NO 130 SEQ ID NO 208 SEQ ID NO 209Lh620 SEQ ID NO 210 SEQ ID NO 211 SEQ ID NO 131 SEQ ID NO 212 SEQ ID NO213 Lh621 SEQ ID NO 214 SEQ ID NO 215 SEQ ID NO 132 SEQ ID NO 216 SEQ IDNO 217 Lh622 SEQ ID NO 218 SEQ ID NO 219 SEQ ID NO 133 SEQ ID NO 220 SEQID NO 221 Lh623 SEQ ID NO 222 SEQ ID NO 223 SEQ ID NO 134 SEQ ID NO 224SEQ ID NO 225 Lh624 SEQ ID NO 226 SEQ ID NO 227 SEQ ID NO 135 SEQ ID NO228 SEQ ID NO 229 Lh625 SEQ ID NO 230 SEQ ID NO 231 SEQ ID NO 136 SEQ IDNO 232 SEQ ID NO 233 Lh626 SEQ ID NO 234 SEQ ID NO 235 SEQ ID NO 137 SEQID NO 236 SEQ ID NO 237 Lh614 SEQ ID NO 238 SEQ ID NO 239 SEQ ID NO 138SEQ ID NO 240 SEQ ID NO 241 Lh627 SEQ ID NO 242 SEQ ID NO 243 SEQ ID NO151 SEQ ID NO 244 SEQ ID NO 245 Lh628 SEQ ID NO 246 SEQ ID NO 247 SEQ IDNO 153 SEQ ID NO 248 SEQ ID NO 249 Lh629 SEQ ID NO 250 SEQ ID NO 251 SEQID NO 155 SEQ ID NO 25 SEQ ID NO 253 Lh630 SEQ ID NO 254 SEQ ID NO 255SEQ ID NO 157 SEQ ID NO 256 SEQ ID NO 257 Lh631 SEQ ID NO 258 SEQ ID NO259 SEQ ID NO 159 SEQ ID NO 260 SEQ ID NO 261 Lh632 SEQ ID NO 262 SEQ IDNO 263 SEQ ID NO 161 SEQ ID NO 264 SEQ ID NO 265 Lh633.2 SEQ ID NO 266SEQ ID NO 267 SEQ ID NO 164 SEQ ID NO 268 SEQ ID NO 269 Lh634.1 SEQ IDNO 270 SEQ ID NO 271 SEQ ID NO 166 SEQ ID NO 272 SEQ ID NO 273 Lh595 SEQID NO 274 SEQ ID NO 275 SEQ ID NO 169 SEQ ID NO 276 SEQ ID NO 277 Lh596SEQ ID NO 278 SEQ ID NO 279 SEQ ID NO 173 SEQ ID NO 280 SEQ ID NO 281

TABLE 7 cDNA sequence (sense strand) Target ID 5′ → 3′ Ld594 SEQ ID NO174 Ld594(b) SEQ ID NO 404 Ld619 SEQ ID NO 176 Ld620 SEQ ID NO 178 Ld583SEQ ID NO 386 Ld584 SEQ ID NO 387 Ld586 SEQ ID NO 388 Ld588 SEQ ID NO389 Ld513 SEQ ID NO 394

TABLE 8 Corresponding amino acid sequence of Target ID cDNA clone asrepresented in Table 9 Ld594 SEQ ID NO 349 Ld594(b) SEQ ID NO 405 Ld619SEQ ID NO 350 Ld620 SEQ ID NO 351 Ld583 SEQ ID NO 390 Ld584 SEQ ID NO391 Ld586 SEQ ID NO 392 Ld588 SEQ ID NO 393 Ld513 SEQ ID NO 395

TABLE 9 Primers Primers dsRNA: sense strand Target Forward Reverserepresented by equivalent ID 5′ → 3′ 5′ → 3′ DNA Sequence 5′ → 3′ Ld594SEQ ID NO 282 SEQ ID NO 283 SEQ ID NO 175 SEQ ID NO 284 SEQ ID NO 285Ld619 SEQ ID NO 286 SEQ ID NO 287 SEQ ID NO 177 SEQ ID NO 288 SEQ ID NO289 Ld620 SEQ ID NO 290 SEQ ID NO 291 SEQ ID NO 179 SEQ ID NO 292 SEQ IDNO 293 Ld513 SEQ ID NO 396 SEQ ID NO 397 SEQ ID NO 400 SEQ ID NO 398 SEQID NO 399

TABLE 10 cDNA Sequence (sense strand) Target ID 5′ → 3′ NI594 SEQ ID NO180 NI619 SEQ ID NO 182 NI626 SEQ ID NO 184 NI537 SEQ ID NO 186

TABLE 11 Corresponding amino acid sequence of cDNA clone as Target IDrepresented in Table 12 NI594 SEQ ID NO 352 NI619 SEQ ID NO 353 NI626SEQ ID NO 354 NI537 SEQ ID NO 355

TABLE 12 Primers Primers dsRNA: sense strand Target Forward Reverserepresented by equivalent ID 5′ → 3′ 5′ → 3′ DNA Sequence 5′ → 3′ NI594SEQ ID NO 294 SEQ ID NO 295 SEQ ID NO 181 SEQ ID NO 296 SEQ ID NO 297NI619 SEQ ID NO 298 SEQ ID NO 299 SEQ ID NO 183 SEQ ID NO 300 SEQ ID NO301 NI626 SEQ ID NO 302 SEQ ID NO 303 SEQ ID NO 185 SEQ ID NO 304 SEQ IDNO 305 NI537 SEQ ID NO 306 SEQ ID NO 307 SEQ ID NO 187 SEQ ID NO 308 SEQID NO 309

TABLE 13 Target Fw primer sequence Reverse primer sequence Ap594 SEQ IDNO 369 SEQ ID NO 370 Ap423 SEQ ID NO 371 SEQ ID NO 372 Ap537 SEQ ID NO373 SEQ ID NO 374 Ap560 SEQ ID NO 375 SEQ ID NO 376

TABLE 14 Target cDNA Sequence (sense strand) ID 5′ → 3′ Ap594 SEQ ID NO188 Ap423 SEQ ID NO 200 Ap537 SEQ ID NO 202 Ap560 SEQ ID NO 204

TABLE 15 Target Corresponding amino acid sequence of ID cDNA clone asrepresented in Table 16 Ap594 SEQ ID NO 356 Ap423 SEQ ID NO 357 Ap537SEQ ID NO 358 Ap560 SEQ ID NO 359

TABLE 16 Primers dsRNA: sense strand Target Primers Forward Reverserepresented by equivalent ID 5′ → 3′ 5′ → 3′ DNA sequence 5′ → 3′ Ap594SEQ ID NO 310 SEQ ID NO 311 SEQ ID NO 189 SEQ ID NO 312 SEQ ID NO 313Ap423 SEQ ID NO 314 SEQ ID NO 315 SEQ ID NO 201 SEQ ID NO 316 SEQ ID NO317 Ap537 SEQ ID NO 318 SEQ ID NO 319 SEQ ID NO 203 SEQ ID NO 320 SEQ IDNO 321 Ap560 SEQ ID NO 322 SEQ ID NO 323 SEQ ID NO 205 SEQ ID NO 324 SEQID NO 325

TABLE 17 Target Forward primer Reverse primer Ld594 SEQ ID NO 377 SEQ IDNO 378

TABLE 18 Target Forward primer Reverse primer NI594 seq id no 379 seq idno 380 NI619 seq id no 381 seq id no 382 NI626 seq id no 383 seq id no384

TABLE 19 Target ID Best Drosophila hit NAME SYMBOL Ld583 CG4759Ribosomal protein L27 RpL27 Ld584 CG 17331 Proteasome, beta-type subunitLd586 CG13704 unknown Ld588 CG4157 Rpn12

TABLE 20 Target ID Best Drosophila hit NAME SYMBOL NI594 CG7178 wings upA wupA (troponin I) NI619 CG7107 troponin T up (upheld) NI626 *CG9073,CG7930, CG2981, troponin C CG12408, CG6514, CG2981, CG7930, CG9073,CG6514, CG12408 NI537 CG32744 Ubiquitin- 5E; protein modificationprocess *unclear: multiple hits in family

TABLE 21 Target ID Best Drosophila hit NAME SYMBOL Ap594 CG7178 wings upA (troponin I) wupA Ap423 CG2746 ribosomal protein L19 RpL19 Ap537CG32744 Ubiquitin-5E; protein modification process Ap560 CG10423ribosomal protein S27 RpS27

It will be appreciated by persons skilled in the art that numerousvariations and/or modifications may be made to the above mentionedassays without departing from the spirit or scope of this assay asgenerically described. Those skilled in the art will recognize, or beable to ascertain using no more than routine experimentation, manyequivalents to the specific examples, and such equivalents are intendedto be encompassed by the present invention. The present example,therefore, is to be considered in all respects as illustrative and notrestrictive.

What is claimed is:
 1. An interfering ribonucleic acid (RNA) thatfunctions upon uptake by an insect pest species to down-regulateexpression of a ribosomal protein S27 target gene in said insect pest,wherein the RNA comprises at least one silencing element, wherein thesilencing element is a region of double-stranded RNA comprising annealedcomplementary strands, one strand of which comprises or consists of asequence of nucleotides which is at least partially complementary to atarget nucleotide sequence within the target gene, and wherein theinterfering RNA is encoded by: (i) a nucleotide sequence comprising anyof SEQ ID NOs 15, 16, or the complement thereof, or a nucleotidesequence that is at least 95% identical to any of SEQ ID NOs 15, 16, orthe complement thereof, (ii) a nucleotide sequence comprising a fragmentof at least 21 contiguous nucleotides of any of SEQ ID NOs 15, 16, thecomplement thereof, (iii) a nucleotide sequence comprising a fragment ofat least 21 contiguous nucleotides of any of SEQ ID NOs 15, 16, or thecomplement thereof, and wherein said nucleotide sequence of saidfragment is at least 95% identical to said corresponding fragment of anyof SEQ ID NOs 15, 16, or the complement thereof, or (iv) a nucleotidesequence comprising a fragment of at least 21 contiguous nucleotides ofa nucleotide sequence encoding an amino acid sequence encoded by SEQ IDNO: 15, 16, the complement thereof.
 2. The interfering RNA of claim 1wherein the silencing element comprises or consists of a sequence of atleast 21 contiguous nucleotides which is complementary or at leastpartially complementary to a target nucleotide sequence within aribosomal protein S27 target gene.
 3. The interfering RNA of claim 1wherein the RNA comprises at least two silencing elements, wherein eachsilencing element comprises or consists of a sequence of nucleotideswhich is at least partially complementary to a target nucleotidesequence within the ribosomal protein S27 target gene.
 4. Theinterfering RNA of claim 3 wherein each of the silencing elementscomprises or consists of a different sequence of nucleotides which iscomplementary to a different target nucleotide sequence within theribosomal protein S27 target gene.
 5. The interfering RNA of claim 4wherein the different ribosomal protein S27 target nucleotide sequencesoriginate from a single ribosomal protein S27 target gene or fromdifferent target genes.
 6. The interfering RNA of claim 5 wherein thedifferent target genes originate from the same insect pest species ordifferent insect pest species.
 7. The interfering RNA of claim 1 whereinthe insect pest species is a plant pest.
 8. The interfering RNA of claim1 wherein the plant pest is an insect pest species selected from theinsect species belonging to the orders Coleoptera, Hemiptera,Lepidoptera, Diptera, Dichyoptera, Orthoptera, and Siphonaptera.
 9. Theinterfering RNA of claim 8 wherein the insect plant pest is selectedfrom the group consisting of Leptinotarsa spp., Nilaparvata spp., Lygusspp., Myzus spp., and Diabrotica spp.
 10. The interfering RNA of claim 1wherein down-regulating expression of the ribosomal protein S27 targetgene causes decreased growth, development, reproduction, or survival ofthe pest as compared with a pest species exposed to an interferingribonucleic acid targeting a non-essential gene or an interferingribonucleic acid that does not down-regulate any genes within the pestspecies.
 11. A polynucleotide comprising a sequence of nucleotidesencoding the interfering RNA of claim
 1. 12. The polynucleotide of claim11 which is comprised in a DNA construct.
 13. The DNA construct of claim12 which is an expression construct, wherein the polynucleotide sequenceencoding the interfering RNA is operably linked to at least oneregulatory sequence capable of driving expression of the polynucleotidesequence.
 14. A host cell comprising the interfering RNA of claim
 1. 15.The host cell of claim 14 wherein the host cell is a prokaryotic or aeukaryotic cell.
 16. The host cell of claim 15 wherein the host cell isa bacterial cell.
 17. A composition for preventing and/or controllinginsect pest infestation comprising the interfering ribonucleic acid(RNA) of claim 1 and at least one suitable carrier, excipient ordiluent.
 18. The composition of claim 17 wherein the interfering RNA isencoded by a polynucleotide comprising a sequence of nucleotidesencoding the interfering RNA which is comprised in a DNA construct,which optionally is an expression construct wherein the polynucleotidesequence encoding the interfering RNA is operably linked to at least oneregulatory sequence capable of driving expression of the polynucleotidesequence.
 19. The composition of claim 17, wherein the interfering RNAis produced by a host cell capable of expressing the interfering RNA.20. The composition of claim 19 wherein the host cell is a bacterialcell.
 21. The composition of claim 17 wherein the composition is in aform suitable for ingestion by an insect.
 22. The composition of claim17 wherein the composition is in solid, liquid or gel form.
 23. Thecomposition of claim 17 wherein the composition is formulated as aninsecticidal spray.
 24. The composition of claim 23 wherein the spray isa pressurized/aerosolized spray or a pump spray.
 25. The composition ofclaim 17 wherein the composition further comprises at least onepesticidal agent selected from the group consisting of a chemicalinsecticide, a patatin, a Bacillus thuringiensis insecticidal protein, aXenorhabdus insecticidal protein, a Photorhabdus insecticidal protein, aBacillus laterosporus insecticidal protein, and a Bacillus sphaericusinsecticidal protein.
 26. The composition of claim 25 wherein saidBacillus thuringiensis insecticidal protein is selected from the groupconsisting of a Cry1, a Cry3, a TIC851, a CryET170, a Cry22, a TIC901, aTIC201, a TIC407, a TIC417, a binary insecticidal protein CryET80 andCryET76, a binary insecticidal protein TIC100 and TIC101, a combinationof an insecticidal protein ET29 or ET37 with an insecticidal proteinTIC810 or TIC812, and a binary insecticidal protein PS149B1.
 27. Ahousing or trap for an insect pest which contains the composition asdefined in claim
 17. 28. A combination for preventing and/or controllingpest infestation comprising the composition of claim 17 and at least oneother active agent.
 29. The combination of claim 28 wherein thecombination is for preventing and/or controlling pest infestation of aplant and the other active agent is an agronomical agent.
 30. Thecombination of claim 29 wherein the agronomical agent comprises aherbicide.
 31. The combination of claim 29 wherein the agronomical agentcomprises a pesticide.
 32. The combination of claim 31 furthercomprising a second pesticide, wherein the second pesticide is selectedfrom the group consisting of a chemical insecticide, a patatin, aBacillus thuringiensis insecticidal protein, a Xenorhabdus insecticidalprotein, a Photorhabdus insecticidal protein, a Bacillus laterosporusinsecticidal protein, and a Bacillus sphaericus insecticidal protein.33. The combination of claim 32 wherein said Bacillus thuringiensisinsecticidal protein is selected from the group consisting of a Cry1, aCry3, a TIC851, a CryET170, a Cry22, a TIC901, a TIC201, a TIC407, aTIC417, a binary insecticidal protein CryET80 and CryET76, a binaryinsecticidal protein TIC100 and TIC101, a combination of an insecticidalprotein ET29 or ET37 with an insecticidal protein TIC810 or TIC812, anda binary insecticidal protein PS149B1.
 34. A method for down-regulatingexpression of a target gene in an insect pest species in order toprevent and/or control pest infestation, comprising contacting said pestspecies with at least one interfering ribonucleic acid (RNA), whereinthe interfering RNA functions upon uptake by the pest to down-regulatethe expression of a target gene within said pest, wherein theinterfering RNA is the interfering RNA of claim
 1. 35. The method ofclaim 34 wherein down-regulation of expression of a target gene in aninsect pest species is used to obtain at least 20% pest control or atleast 20% pest mortality as compared to control insect pests contactedwith an interfering ribonucleic acid (RNA) targeting a non-essentialpest gene or a target gene not expressed in said pest.
 36. The method ofclaim 34 wherein the method is used to prevent and/or control pestinfestation of a plant.
 37. The method of claim 36 wherein the plant isselected from the group consisting of cotton, potato, rice, canola,sunflower, sorghum, pearl millet, corn, strawberries, soy, alfalfa,tomato, eggplant, pepper and tobacco.
 38. A kit comprising theinterfering ribonucleic acid (RNA) of claim 1, the DNA construct ofclaim 12, or the composition of claim 17 for preventing and/orcontrolling insect pest infestation.